CN106619718A - Cellular preparation and preparation method and application thereof - Google Patents
Cellular preparation and preparation method and application thereof Download PDFInfo
- Publication number
- CN106619718A CN106619718A CN201610906738.3A CN201610906738A CN106619718A CN 106619718 A CN106619718 A CN 106619718A CN 201610906738 A CN201610906738 A CN 201610906738A CN 106619718 A CN106619718 A CN 106619718A
- Authority
- CN
- China
- Prior art keywords
- preparation
- cell
- cell preparation
- stem cells
- mesenchymal stem
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 69
- 230000001413 cellular effect Effects 0.000 title abstract 5
- 210000004623 platelet-rich plasma Anatomy 0.000 claims abstract description 20
- 210000002901 mesenchymal stem cell Anatomy 0.000 claims abstract description 17
- 206010019280 Heart failures Diseases 0.000 claims abstract description 16
- 210000004027 cell Anatomy 0.000 claims description 54
- 210000000130 stem cell Anatomy 0.000 claims description 28
- 210000004271 bone marrow stromal cell Anatomy 0.000 claims description 16
- 238000011282 treatment Methods 0.000 claims description 14
- 239000003814 drug Substances 0.000 claims description 12
- 229940079593 drug Drugs 0.000 claims description 4
- 210000005259 peripheral blood Anatomy 0.000 claims description 4
- 239000011886 peripheral blood Substances 0.000 claims description 4
- 210000003716 mesoderm Anatomy 0.000 claims description 3
- 239000002671 adjuvant Substances 0.000 claims description 2
- 238000002483 medication Methods 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 8
- 241000699670 Mus sp. Species 0.000 description 8
- 239000006285 cell suspension Substances 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 206010007556 Cardiac failure acute Diseases 0.000 description 4
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 4
- 102000013275 Somatomedins Human genes 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 210000002249 digestive system Anatomy 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 3
- 239000002158 endotoxin Substances 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 102000009024 Epidermal Growth Factor Human genes 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 description 2
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 description 2
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 2
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000000747 cardiac effect Effects 0.000 description 2
- 206010007625 cardiogenic shock Diseases 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 210000003141 lower extremity Anatomy 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- BQJCRHHNABKAKU-KBQPJGBKSA-N morphine Chemical compound O([C@H]1[C@H](C=C[C@H]23)O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4O BQJCRHHNABKAKU-KBQPJGBKSA-N 0.000 description 2
- 230000002685 pulmonary effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 2
- 208000010444 Acidosis Diseases 0.000 description 1
- 206010001029 Acute pulmonary oedema Diseases 0.000 description 1
- 206010007558 Cardiac failure chronic Diseases 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 206010013954 Dysphoria Diseases 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 101800003838 Epidermal growth factor Proteins 0.000 description 1
- 208000001953 Hypotension Diseases 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 206010027417 Metabolic acidosis Diseases 0.000 description 1
- 108020001621 Natriuretic Peptide Proteins 0.000 description 1
- 102000004571 Natriuretic peptide Human genes 0.000 description 1
- 206010031123 Orthopnoea Diseases 0.000 description 1
- 208000037656 Respiratory Sounds Diseases 0.000 description 1
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 1
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 1
- 206010047924 Wheezing Diseases 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000002876 beta blocker Substances 0.000 description 1
- 229940097320 beta blocking agent Drugs 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000008828 contractile function Effects 0.000 description 1
- 238000011443 conventional therapy Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000003205 diastolic effect Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 231100000284 endotoxic Toxicity 0.000 description 1
- 230000002346 endotoxic effect Effects 0.000 description 1
- 229940116977 epidermal growth factor Drugs 0.000 description 1
- 239000003172 expectorant agent Substances 0.000 description 1
- 230000003419 expectorant effect Effects 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 230000000004 hemodynamic effect Effects 0.000 description 1
- 230000001951 hemoperfusion Effects 0.000 description 1
- 230000036543 hypotension Effects 0.000 description 1
- 208000018875 hypoxemia Diseases 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 239000004041 inotropic agent Substances 0.000 description 1
- 239000002171 loop diuretic Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000005399 mechanical ventilation Methods 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 229960005181 morphine Drugs 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 239000000692 natriuretic peptide Substances 0.000 description 1
- 238000013546 non-drug therapy Methods 0.000 description 1
- 208000012144 orthopnea Diseases 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 238000010837 poor prognosis Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 206010037833 rales Diseases 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000036391 respiratory frequency Effects 0.000 description 1
- 230000033764 rhythmic process Effects 0.000 description 1
- 238000007665 sagging Methods 0.000 description 1
- 238000011477 surgical intervention Methods 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 230000035488 systolic blood pressure Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 239000003634 thrombocyte concentrate Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 230000001256 tonic effect Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 1
- 239000005526 vasoconstrictor agent Substances 0.000 description 1
- 229940124549 vasodilator Drugs 0.000 description 1
- 239000003071 vasodilator agent Substances 0.000 description 1
- 230000002861 ventricular Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/16—Blood plasma; Blood serum
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Cell Biology (AREA)
- Developmental Biology & Embryology (AREA)
- Hematology (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Virology (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention relates to the technical field of cellular preparations, in particular to a cellular preparation and a preparation method and application thereof. The cellular preparation is platelet-rich plasma containing mesenchymal stem cells. The study of the invention discovers that the joint use of mesenchymal stem cells and PRP (platelet-rich plasma) provides significant increase in LVEF level and significant decrease in BNP level, with the effect better than that of the sole use of mesenchymal stem cells or PRP, and the effective rate is up to 80-90%. It can be seen that the cellular preparation is effective in treating heart failure.
Description
Technical field
The present invention relates to cell preparation technical field, more particularly to a kind of cell preparation and its preparation method and application.
Background technology
Heart failure (heart failure) abbreviation heart failure, refers to the contractile function and (or) diastolic function due to heart
Generation obstacle, it is impossible to which venous return is fully discharged heart, causes Venous system sludging, Arterial system hemoperfusion
Deficiency, so as to cause cardiac cycle obstacle syndrome, shows as pulmonary venous pleonaemia, caval vein congestion in this kind of obstacle disease cluster.Root
According to the emergency that heart failure occurs, clinic can be divided into acute heart failure and chronic heart failure.Acute heart failure can cause acute
Pulmonary edema, onset urgency, the state of an illness can be developed rapidly to critical state.The severe dyspnea of burst, orthopnea, gasp for breath,
Dysphoria simultaneously has fear, and respiratory frequency is up to 30～50 beats/min；Frequently cough simultaneously brings up a large amount of pink foam sample expectorant；
Heart rate is fast, apex Chang Kewen and gallop rhythm；Two fullness of the lung cloth moist rales and wheezing sound.Cardiogenic shock can also be caused, shown as
Hypotension, tissue low perfusion state, hemodynamics obstacle, metabolic acidosiss and hypoxemia.Acute heart failure often jeopardizes life
Life, it is necessary to urgent rescue.
Current clinical treatment is mainly using means such as Drug therapy, interventional therapy and surgical interventions：(1) initial therapy is
Jing masks or nasal tube oxygen inhalation；The Jing veins such as morphine, loop diuretic, cardiac tonic give.Patient is set to take seat or semireclining position, two lower limbs
It is sagging, reduce veins of lower extremity backflow.(2) still not alleviation person should select to be lived using blood vessel the state of an illness according to systolic pressure and pulmonary venous pleonaemia situation
Property medicine, such as inotropic agent, vasodilator and vasoconstrictor.(3) be in a bad way, blood pressure persistently reduce (<
90mmHg) or even cardiogenic shock person, monitor blood flow kinetics are answered, and supported using intra aortic balloon counterpulsation, mechanical ventilation,
The various non-drug therapy methods such as blood purification, ventricle assist and surgical operation.(4) dynamic measurement BNP/NT-
ProBNP contributes to instructing the treatment of acute heart failure, and its level still can be in any more person after treatment, points out poor prognosis, should strengthen controlling
Treat；Its level is reduced and the range of decrease after treatment>30%, point out treatment effective, good prognosis.But these Therapeutic Method can only be in certain journey
Alleviate the generation of heart failure on degree.
Stem cell is the pluripotent cell that a class has the of self-replication capacity and differentiation capability, with the various organizers of regeneration
Official and the potential function of human body, medical circle is referred to as " general-purpose cell ".Find that medulla mesenchyma is done through the research of forefathers
Cell directly or indirectly can be repaiied as a kind of medicine under suitable Cytokine to impaired myocardial cell
It is multiple or supplementary, one of study hotspot of heart failure treatment is had become at present.Therefore, research and develop it is a kind of can effectively treatment heart failure it is dry
Cell preparation becomes inexorable trend.
The content of the invention
In view of this, the invention provides a kind of cell preparation and its preparation method and application.The cell preparation can be effective
Treatment heart failure.
In order to realize foregoing invention purpose, the present invention provides technical scheme below：
The invention provides a kind of cell preparation, it includes the platelet rich plasma of mescenchymal stem cell.
Platelet rich plasma (Platelet-rich plasma, PRP) refers to the blood that autologous whole blood is obtained Jing after centrifugation
Platelet concentrate, containing the raised growth factor and protein.Contain substantial amounts of somatomedin, such as epidermal growth factor in platelet
(EGF), platelet derived growth factor (PDGF), transforming growth factor-β (TGF-β), insulin like growth factor (IGF), blood
Endothelial tube somatomedin (VEGF) etc..Because it plays highly important effect in terms of tissue regeneration, in recent years, it is facing
Bed is especially valued by the people in the wound and defect repair in oral cavity, and achieves the effect for making us more being satisfied with.
The present invention can preferably play the reparation of mescenchymal stem cell by after PRP and mesenchyma stem cell combined use
Effect, so as to be remarkably improved left ventricular ejection fraction (LVEF) level, and can significantly reduce natriuretic peptide (BNP) level, effect
It is better than exclusive use mescenchymal stem cell or PRP, effective percentage is up to 80%～90%.It can be seen that, cell preparation of the present invention can be effective
Treatment heart failure.
Preferably, (1～10) × 10 containing mescenchymal stem cell in per mL platelet rich plasmas6It is individual.
In the embodiment that the present invention is provided, in every mL platelet rich plasmas mescenchymal stem cell 1 × 10 is contained6It is individual.
In another embodiment that the present invention is provided, in every mL platelet rich plasmas mescenchymal stem cell 5 × 10 is contained6It is individual.
In another embodiment that the present invention is provided, in every mL platelet rich plasmas mescenchymal stem cell 1 × 10 is contained7It is individual.
In the embodiment that the present invention is provided, mescenchymal stem cell is mesenchymal stem cells MSCs.
Preferably, mesenchymal stem cells MSCs be P3 for mesenchymal stem cells MSCs.
Present invention also offers the preparation method of the cell preparation, comprises the steps：
Using the resuspended mescenchymal stem cell of platelet rich plasma, cell preparation is obtained.
In the present invention, the preparation method of mescenchymal stem cell is：The culture medium comprising mescenchymal stem cell is taken, training is removed
Foster base, is digested using Digestive system, obtains single cell suspension；Single cell suspension is centrifuged, is washed, obtained mesenchyme and do
Cell.
In the present invention, peripheral blood is centrifuged, obtains platelet rich plasma.
In the embodiment that the present invention is provided, the preparation method of platelet rich plasma is specially：Peripheral blood is adopted into 300g
Rotating speed is centrifuged 5min, then 5min is centrifuged using 300g rotating speeds, finally 5min is centrifuged using 800g rotating speeds.
Present invention also offers the cell preparation is preparing the application in improving LVEF levels or the horizontal medicines of reduction BNP.
Present invention also offers application of the cell preparation in treatment heart failure medications are prepared.
Present invention also offers a kind of medicine, including the cell preparation that the present invention is provided.The cell preparation is comprising filling
The platelet rich plasma of matter stem cell.
In the embodiment that the present invention is provided, medicine also includes pharmaceutically acceptable adjuvant.
The invention provides a kind of cell preparation and its preparation method and application.The cell preparation is to include that mesenchyme is dry thin
The platelet rich plasma of born of the same parents.The present invention has the advantages that：
Present invention research discovery, after mescenchymal stem cell and PRP are used in combination, is remarkably improved LVEF levels, and can
BNP levels are significantly reduced, effect is better than exclusive use mescenchymal stem cell or PRP, and effective percentage is up to 80%～90%.It can be seen that,
Cell preparation of the present invention can effectively treatment heart failure.
Specific embodiment
The invention discloses a kind of cell preparation and its preparation method and application, those skilled in the art can use for reference herein
Content, is suitably modified technological parameter realization.Specifically, all similar replacements and change are to people in the art
It is it will be apparent that they are considered as being included in the present invention for member.The method of the present invention and application have passed through preferably real
Apply example to be described, related personnel substantially can be in without departing from present invention, spirit and scope to method described herein
It is modified with application or suitably the technology of the present invention is realized and applied to change with combining.
Biomaterial used can be buied by market in cell preparation that the present invention is provided and its preparation method and application.
With reference to embodiment, the present invention is expanded on further：
Embodiment 1：The preparation of human marrow mesenchymal stem cell
Bone marrow 10mL is obtained from regular purchasing channel, Percoll (1.073 × 10 after anticoagulant heparin-3G/L) density gradient from
The heart, is taken middle mononuclear cell layer and is washed 2 times with low sugar DMEM, is inoculated with into 75cm2Culture bottle, with low sugar DMEM containing 10%FBS
Culture.
Liquid is changed after 24h, not adherent cell is abandoned, remaining attached cell is MSCs.
Liquid was changed per 2～3 days, 0.25% trypsinization is passed on after cell growth is converged to 90%.
Embodiment 2：The preparation of PRP
Peripheral blood in patients 10mL, 300g centrifugation 5min are taken, then again 300g is centrifuged 5min, last 800g centrifugations
5min, obtains PRP.
Embodiment 3：The preparation of cell preparation
P3 prepared by Example 1 sucks culture medium for mesenchymal stem cells MSCs, adds Digestive system to be digested, it
Terminated afterwards digesting with appropriate serum, gently blow and beat into single cell suspension.1500rpm, is centrifuged 5min, abandons supernatant, is washed with 4 DEG C of PBS
Resuspended mixing after cell 3 times is washed, adjustment cell concentration is about 107Individual/mL.
Using the resuspended P3 of PRP of the acquisition of embodiment 2 for mesenchymal stem cells MSCs, cell preparation is obtained, its composition is as follows
Shown in table.
The cell preparation of 1 embodiment of table 3 is constituted
| Component | Content |
| P3 is for mesenchymal stem cells MSCs | 1×106Individual/mL |
| PRP | 3.5mL |
Embodiment 4：The preparation of cell preparation
P3 prepared by Example 1 sucks culture medium for mesenchymal stem cells MSCs, adds Digestive system to be digested, it
Terminated afterwards digesting with appropriate serum, gently blow and beat into single cell suspension.1500rpm, is centrifuged 5min, abandons supernatant, is washed with 4 DEG C of PBS
Resuspended mixing after cell 3 times is washed, adjustment cell concentration is about 107Individual/mL.
Using the resuspended P3 of PRP of the acquisition of embodiment 2 for mesenchymal stem cells MSCs, cell preparation is obtained, its composition is as follows
Shown in table.
The cell preparation of 2 embodiment of table 4 is constituted
| Component | Content |
| P3 is for mesenchymal stem cells MSCs | 5×106Individual/mL |
| PRP | 3.5mL |
Embodiment 5：The preparation of cell preparation
P3 prepared by Example 1 sucks culture medium for mesenchymal stem cells MSCs, adds Digestive system to be digested, it
Terminated afterwards digesting with appropriate serum, gently blow and beat into single cell suspension.1500rpm, is centrifuged 5min, abandons supernatant, is washed with 4 DEG C of PBS
Resuspended mixing after cell 3 times is washed, adjustment cell concentration is about 107Individual/mL.
Using the resuspended P3 of PRP of the acquisition of embodiment 2 for mesenchymal stem cells MSCs, cell preparation is obtained, its composition is as follows
Shown in table.
The cell preparation of 3 embodiment of table 5 is constituted
| Component | Content |
| P3 is for mesenchymal stem cells MSCs | 1×107Individual/mL |
| PRP | 3.5mL |
Embodiment 6：Cell preparation is evaluated
Microorganism detection and endotoxin detection are carried out to cell preparation obtained in embodiment 3-5.Wherein, microorganism detection
(antibacterial, funguses) are detected according to the 4th general rule 1100 of version Pharmacopoeia of People's Republic of China in 2015；Endotoxin detects basis
The 4th general rule 1143 of version Pharmacopoeia of People's Republic of China in 2015 is detected.
Testing result：Biological preparation obtained in embodiment 3-5 is negative in antibacterial, funguses and endotoxic detection,
Endotoxin report shows<0.25EU/mL.The up-to-standard of the stem cell medicine is illustrated, can be used for the reparation of wound.
Embodiment 7：The repairing effect detection of preparation
Test method：Using intramyocardial injection mode, preparation injection is carried out to the successful mice of heart failure modeling.
Take the successful mice of 60 heart failure modelings, be divided into 6 groups, including negative control group, 3 groups of experimental grouies, stem cell group and
PRP groups：
Negative control group：Using beta-blocker conventional therapy means；
Experimental group：Treated with preparation obtained in embodiment 3-5 respectively；
Stem cell group：The stem cell obtained using embodiment 1 is treated；
PRP groups：The PRP obtained using embodiment 2 is treated.
Each group is as follows to the injection dosage of mice：
Injection dosage of each group of table 4 to mice
| Group | Mice quantity (only) | Dosage |
| Negative control group | 28 | —— |
| Embodiment 3 | 28 | 1mL |
| Embodiment 4 | 28 | 1mL |
| Embodiment 5 | 28 | 1mL |
| Stem cell group | 28 | 2×107It is individual |
| PRP groups | 28 | 2mL |
The indexs such as the LVEF and BNP levels of mice are observed after 8 weeks, the effective percentage for the treatment of is passed judgment on.
Effective percentage calculation is：Mice quantity/28 that LVEF and BNP indexs are significantly improved.
As a result it is as shown in the table：
The indexs such as the LVEF and BNP levels of the mice of table 5
Note：* represent compared with matched group with significant difference, p<0.05.
From above-mentioned result of the test, after mescenchymal stem cell and PRP are used in combination, LVEF levels are remarkably improved,
And BNP levels can be significantly reduced, effect is better than exclusive use mescenchymal stem cell or PRP, and effective percentage is up to 80%～90%.Can
See, cell preparation of the present invention can effectively treatment heart failure.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of cell preparation, it is characterised in that the cell preparation is the platelet rich plasma comprising mescenchymal stem cell.
2. cell preparation according to claim 1, it is characterised in that contain mescenchymal stem cell in per mL platelet rich plasmas
(1～10) × 106It is individual.
3. cell preparation according to claim 1 and 2, it is characterised in that the mescenchymal stem cell is medulla mesenchyma
Stem cell.
4. cell preparation according to any one of claim 1 to 3, it is characterised in that the mesenchymal stem cells MSCs
It is P3 for mesenchymal stem cells MSCs.
5. as any one of Claims 1-4 cell preparation preparation method, it is characterised in that comprise the steps：
Using the resuspended mescenchymal stem cell of platelet rich plasma, the cell preparation is obtained.
6. preparation method according to claim 5, it is characterised in that the preparation method of the platelet rich plasma is concrete
For：Peripheral blood is centrifuged into 5min using 300g rotating speeds, then 5min is centrifuged using 300g rotating speeds, be finally centrifuged using 800g rotating speeds
5min。
7. cell preparation is preparing raising LVEF levels or is reducing in the horizontal medicines of BNP as any one of Claims 1-4
Application.
8. application of the cell preparation in treatment heart failure medications are prepared as any one of Claims 1-4.
9. a kind of medicine, it is characterised in that include the cell preparation as any one of Claims 1-4.
10. medicine according to claim 9, it is characterised in that the medicine also includes pharmaceutically acceptable adjuvant.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610906738.3A CN106619718A (en) | 2016-10-18 | 2016-10-18 | Cellular preparation and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610906738.3A CN106619718A (en) | 2016-10-18 | 2016-10-18 | Cellular preparation and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106619718A true CN106619718A (en) | 2017-05-10 |
Family
ID=58855916
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610906738.3A Pending CN106619718A (en) | 2016-10-18 | 2016-10-18 | Cellular preparation and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106619718A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110585243A (en) * | 2019-08-12 | 2019-12-20 | 丰泽康生物医药（深圳）有限公司 | Pluripotent cell active matter and platelet-rich plasma compound for treating glucocorticoid-dependent dermatitis as well as preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102438636A (en) * | 2009-05-20 | 2012-05-02 | 卡迪欧参生物科技有限公司 | Pharmaceutical composition for treating heart diseases |
| CN103860597A (en) * | 2014-03-03 | 2014-06-18 | 奥思达干细胞有限公司 | Stem cell preparation for treating ischemic cardiomyopathy and preparation method of stem cell preparation |
-
2016
- 2016-10-18 CN CN201610906738.3A patent/CN106619718A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102438636A (en) * | 2009-05-20 | 2012-05-02 | 卡迪欧参生物科技有限公司 | Pharmaceutical composition for treating heart diseases |
| CN103860597A (en) * | 2014-03-03 | 2014-06-18 | 奥思达干细胞有限公司 | Stem cell preparation for treating ischemic cardiomyopathy and preparation method of stem cell preparation |
Non-Patent Citations (2)
| Title |
|---|
| K.YAMAHARA等: "Pathological role of angiostatin in heart failure: an endogenous inhibitor of mesenchymal stem-cell activation", 《HEART》 * |
| ZHIHUA FANG等: "Functional characterization of human umbilical cord-derived mesenchymal stem cells for treatment of systolic heart failure", 《EXP.THER.MED.》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110585243A (en) * | 2019-08-12 | 2019-12-20 | 丰泽康生物医药（深圳）有限公司 | Pluripotent cell active matter and platelet-rich plasma compound for treating glucocorticoid-dependent dermatitis as well as preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Strauer et al. | Stem cell therapy in perspective | |
| JP2010155842A5 (en) | ||
| Kang et al. | Aged human cells rejuvenated by cytokine enhancement of biomaterials for surgical ventricular restoration | |
| JP2010523264A5 (en) | ||
| EP2968409A1 (en) | Treatment of inflammatory respiratory disease using biological solutions | |
| CN110193027A (en) | A kind of preparation method and new opplication of stem cell excretion body | |
| Rees et al. | Cardiac resuscitation with percutaneous cardiopulmonary support | |
| Wang et al. | Implantable and biodegradable macroporous iron oxide frameworks for efficient regeneration and repair of infracted heart | |
| Wu et al. | Ivabradine promotes angiogenesis and reduces cardiac hypertrophy in mice with myocardial infarction. | |
| CN104862281A (en) | Mesenchymal stem cell modified by recombinant vector and used for expressing bFGF and PDGF-BB as well as preparation method and application of mesenchymal stem cell | |
| CN106619718A (en) | Cellular preparation and preparation method and application thereof | |
| CN110462023A (en) | Composition for promoting stem cell activity comprising histone deacetylase inhibitor and activator as active ingredients | |
| CN105816513A (en) | Medicine for treating coronary microvascular dysfunction and preparation method thereof | |
| Plácido et al. | Nonpharmacological management of acute heart failure | |
| US20250002580A1 (en) | Method for inhibiting cardiac fibroblast transdifferentiation | |
| Tan et al. | Biomaterial‐Facilitated Local Delivery of Stem Cell‐Derived Small Extracellular Vesicles: Perspectives in Surgical Therapy | |
| CN106474456A (en) | A kind of cell preparation and its preparation method and application | |
| CN108904782A (en) | CTRP3 is used to prepare the application of prophylactic treatment myocardial hypertrophy drug | |
| Hayashi et al. | Intervening to preserve function in ischemic cardiomyopathy with a porous hydrogel and extracellular matrix composite in a rat myocardial infarction model | |
| KR101555941B1 (en) | Process for Enhancing Stem Cell Bioactivity Using Fucoidan, and The Cellular Therapeutic Supplementary Agent Comprising The Same | |
| JP3977452B2 (en) | Arterial disease treatment | |
| Huang et al. | Effect of bFGF gel nano-sustained-release technology on rehabilitation effect of adult heart valve replacement | |
| US20190255117A1 (en) | Synoviolin expression inhibitor containing mesenchymal stem cell or culture supernatant thereof | |
| Zhang et al. | Sequential and timely transfection of hepatocyte growth factor and monocyte chemotactic protein-1 ameliorates hyperkinetic pulmonary artery hypertension in rabbits | |
| JP6383938B2 (en) | Treatment of glottic insufficiency with fat containing high-concentration adipose tissue-derived mesenchymal stem cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170510 |
|
| RJ01 | Rejection of invention patent application after publication |