CN101045915A - Oocyte germinal vecicle transplanting technology - Google Patents
Oocyte germinal vecicle transplanting technology Download PDFInfo
- Publication number
- CN101045915A CN101045915A CNA2006100709884A CN200610070988A CN101045915A CN 101045915 A CN101045915 A CN 101045915A CN A2006100709884 A CNA2006100709884 A CN A2006100709884A CN 200610070988 A CN200610070988 A CN 200610070988A CN 101045915 A CN101045915 A CN 101045915A
- Authority
- CN
- China
- Prior art keywords
- germinal vesicle
- ovocyte
- technology
- zona pellucida
- nuclear transfer
- 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
- 210000000287 oocyte Anatomy 0.000 title claims abstract description 26
- 210000004340 zona pellucida Anatomy 0.000 claims abstract description 16
- 235000015097 nutrients Nutrition 0.000 claims description 4
- 108010059712 Pronase Proteins 0.000 claims description 3
- CJGYSWNGNKCJSB-YVLZZHOMSA-N bucladesine Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](OC(=O)CCC)[C@@H]2N1C(N=CN=C2NC(=O)CCC)=C2N=C1 CJGYSWNGNKCJSB-YVLZZHOMSA-N 0.000 claims description 3
- 229960005263 bucladesine Drugs 0.000 claims description 3
- UBDHSURDYAETAL-UHFFFAOYSA-N 8-aminonaphthalene-1,3,6-trisulfonic acid Chemical compound OS(=O)(=O)C1=CC(S(O)(=O)=O)=C2C(N)=CC(S(O)(=O)=O)=CC2=C1 UBDHSURDYAETAL-UHFFFAOYSA-N 0.000 claims description 2
- XXPDBLUZJRXNNZ-UHFFFAOYSA-N promethazine hydrochloride Chemical compound Cl.C1=CC=C2N(CC(C)N(C)C)C3=CC=CC=C3SC2=C1 XXPDBLUZJRXNNZ-UHFFFAOYSA-N 0.000 claims description 2
- GBOGMAARMMDZGR-UHFFFAOYSA-N UNPD149280 Natural products N1C(=O)C23OC(=O)C=CC(O)CCCC(C)CC=CC3C(O)C(=C)C(C)C2C1CC1=CC=CC=C1 GBOGMAARMMDZGR-UHFFFAOYSA-N 0.000 claims 2
- GBOGMAARMMDZGR-JREHFAHYSA-N cytochalasin B Natural products C[C@H]1CCC[C@@H](O)C=CC(=O)O[C@@]23[C@H](C=CC1)[C@H](O)C(=C)[C@@H](C)[C@@H]2[C@H](Cc4ccccc4)NC3=O GBOGMAARMMDZGR-JREHFAHYSA-N 0.000 claims 2
- GBOGMAARMMDZGR-TYHYBEHESA-N cytochalasin B Chemical compound C([C@H]1[C@@H]2[C@@H](C([C@@H](O)[C@@H]3/C=C/C[C@H](C)CCC[C@@H](O)/C=C/C(=O)O[C@@]23C(=O)N1)=C)C)C1=CC=CC=C1 GBOGMAARMMDZGR-TYHYBEHESA-N 0.000 claims 2
- 239000007788 liquid Substances 0.000 claims 1
- 238000012546 transfer Methods 0.000 description 22
- 102000002322 Egg Proteins Human genes 0.000 description 11
- 108010000912 Egg Proteins Proteins 0.000 description 11
- 210000004681 ovum Anatomy 0.000 description 11
- 238000000034 method Methods 0.000 description 7
- 230000008569 process Effects 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 230000004927 fusion Effects 0.000 description 3
- 230000035800 maturation Effects 0.000 description 3
- CWHJIJJSDGEHNS-MYLFLSLOSA-N Senegenin Chemical compound C1[C@H](O)[C@H](O)[C@@](C)(C(O)=O)[C@@H]2CC[C@@]3(C)C(CC[C@]4(CCC(C[C@H]44)(C)C)C(O)=O)=C4[C@@H](CCl)C[C@@H]3[C@]21C CWHJIJJSDGEHNS-MYLFLSLOSA-N 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000035558 fertility Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 210000004508 polar body Anatomy 0.000 description 2
- 239000009871 tenuigenin Substances 0.000 description 2
- 241000863032 Trieres Species 0.000 description 1
- 208000036878 aneuploidy Diseases 0.000 description 1
- 231100001075 aneuploidy Toxicity 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- JVHIPYJQMFNCEK-UHFFFAOYSA-N cytochalasin Natural products N1C(=O)C2(C(C=CC(C)CC(C)CC=C3)OC(C)=O)C3C(O)C(=C)C(C)C2C1CC1=CC=CC=C1 JVHIPYJQMFNCEK-UHFFFAOYSA-N 0.000 description 1
- ZMAODHOXRBLOQO-UHFFFAOYSA-N cytochalasin-A Natural products N1C(=O)C23OC(=O)C=CC(=O)CCCC(C)CC=CC3C(O)C(=C)C(C)C2C1CC1=CC=CC=C1 ZMAODHOXRBLOQO-UHFFFAOYSA-N 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention is technology of transplanting oocyte germinal vesicle with zona pellucida eliminated, and belongs to the field of bioengineering technology. The present invention eliminates zona pellucida from oocyte to facilitate microscope operation. The technology of the present invention can reach the aim of transplanting oocyte germinal vesicle and make germinal vesicle transplanting high in efficiency, low in cost and simple.
Description
The invention belongs to biotechnology, particularly the experiment research of biology and medical science and clinical application.
Germinal vesicle nuclear transfer is a kind of technology of just setting up recent years, this technology is widely used in setting up interactional cell model between nucleus and the tenuigenin, and research, the especially latter and the human reproduction's health of getting in touch etc. further inquired between ovocyte matter and the ovocyte dysploidy are closely bound up.Utilize the germinal vesicle nuclear transfer technology can reduce women's at advanced age (more than 40 years old) fertility risk, reach the purpose of prenatal and postnatal care.Germinal vesicle nuclear transfer mainly is meant and takes out the cytosome (Cytoplast) (this cytosome comprises the GV phase ovocyte of stoning, MII phase ovocyte and zygote) of the nucleome (Karyoplast) that forms and stoning gained by micrurgy and merge the process that forms recombinant eggs being in the germinal vesicle of ovocyte in germinal vesicle period and small amounts of cells matter.The present invention further simplifies the germinal vesicle nuclear transfer technology, utilizes zona pellucida to remove combining of art and germinal vesicle nuclear transfer art, makes the germinal vesicle nuclear transfer technology that previous efficient is low, cost is high, difficulty is big that new breakthrough arranged.The learning time that can make the tyro is from the time decreased to 1 of previous some months day.
The objective of the invention is: utilize zona pellucida to remove art and combine, make that the situation that the germinal vesicle nuclear transfer technology is low from previous efficient, cost is high, difficulty is big is converted into efficiently, low-cost, easy novel germinal vesicle nuclear transfer technology with the germinal vesicle nuclear transfer technology.The learning time that particularly can make the tyro is from the time decreased to 1 of previous some months day.
Content of the present invention is: the disadvantage of current germinal vesicle nuclear transfer technology is exactly that efficient is low, and the cost height is high to trier's state of the art requirement.We adopt zona pellucida to remove art and make ovocyte be more convenient for operating in whole process, reduced operating process, have reduced operation easier, have improved working efficiency.Zona pellucida is removed art apply to germinal vesicle nuclear transfer, make this technology become efficient, with low cost, experimenter's technical background is not had specific requirement.Thereby make the germinal vesicle nuclear transfer technology need highly difficult technical change that the professional person operates for simply being applicable to general personnel's routine techniques by original.
(1) uses the PRONASE A of 0.1-1.0% or the zona pellucida that ovocyte is removed in T acid.
(2) will go the ovocyte of zona pellucida in the nutrient solution that contains 5 μ g/ml-25 μ g/ml cytochalasin Bs and IBMX or dbcAMP, to handle for some time, carry out micrurgy to take out germinal vesicle producing the ovocyte that recovers again after the distortion.
(3) germinal vesicle that will remove the cytosome of germinal vesicle and consubstantiality or allosome carries out bondingly in the nutrient solution of the PHA that contains 10 μ g/ml-100 μ g/ml, then carries out electricity and merges.
(4) select the reconstruct ovum of fusion fully to carry out the maturation in vitro cultivation, observe polar body after 12 hours and discharge situation.
(5) ovocyte that will discharge polar body carries out the micrurgy second time to obtain the nuclear district, and this is examined the district carry out bonding and electric fusion with the cytosome of the MII phase ovocyte of removing the nuclear district of cylinder mature, reconstruct ovum after the fusion activates processing external, is transferred to nutrient solution subsequently and grows cultivation.The embryo in resulting each period can select transplanting or biology of being correlated with or medical experiment research in the organism
Advantage of the present invention and positively effect
1, the present invention is because the zona pellucida of the ovum that adopts all is removed, so do not need to carry out special zona oepning, ovum need not carry out 2 hours cultivation to wait for the formation in ovum week crack external yet simultaneously.And current current germinal vesicle nuclear transfer technology must be with ovum in vitro culture more than at least 2 hours, has only the opening that occurs could being undertaken by county's micro-operation during the crack in ovum week zona pellucida when ovocyte.Therefore, the present invention has not only shortened experimental period but also has reduced the bigger micrurgy of a step difficulty, has improved conventional efficient.
2, the ovum pin of holding used in the present invention is held the ovum pin for remaining silent, and can make operator not need control to hold the ovum pin like this.Current current germinal vesicle nuclear transfer technical requirements operator control simultaneously and hold ovum pin and kernel removing needle.Therefore, the present invention has simplified operating process, has reduced operation easier, has improved operation efficiency.
3, the germinal vesicle nuclear transfer technology comprises the micrurgy of two different times.Therefore, the present invention has reduced operation easier on the whole, has improved working efficiency.
4, utilize the zona pellucida ovocyte germinal vesicle nuclear transfer technology of going, can so that the germinal vesicle nuclear transfer technology from before poor efficiency, expensively be converted into efficiently, new technique scheme cheaply.So just improved the popularization of ovocyte germinal vesicle nuclear transfer technology widely, thereby be relevant research, for example: the generation of the chromosomal aneuploidy that mutual relationship and advanced age are occurred in women's oocyte maturation process between the nucleus of oocyte maturation and the tenuigenin and advanced age the women artificial fertility established solid basis.
5, utilize ovocyte to go zona pellucida germinal vesicle nuclear transfer technology, new research thinking can also be provided for nuclear transfer technology from now on, promptly in conjunction with the micrurgy of going the zona pellucida technology.
In sum, go zona pellucida ovocyte germinal vesicle nuclear transfer technology as a new technology have easy to learn, save time, laborsaving, good effectiveness.
Claims (6)
1, use PRONASE A or T acid to remove the zona pellucida of ovocyte.
2, the concentration of pronase is 0.1-1.0% according to claim 1.
3, the ovocyte that will remove zona pellucida is handled for some time in the operation liquid of cytochalasin B and IBMX or dbcAMP, carries out micrurgy to producing the ovocyte that recovers again after the distortion.
4, the concentration as cytochalasin B as described in the claim 3 is 5 μ g/ml-50 μ g/ml, and the concentration of IBMX or dbcAMP is 10-100 μ g/ml.
5, will remove the cytosome of germinal vesicle and the germinal vesicle of consubstantiality or allosome carries out bonding in containing the nutrient solution of PHA.
6, the concentration as PHA as described in the claim 5 is 10 μ g/ml-100 μ g/ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006100709884A CN101045915A (en) | 2006-03-29 | 2006-03-29 | Oocyte germinal vecicle transplanting technology |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006100709884A CN101045915A (en) | 2006-03-29 | 2006-03-29 | Oocyte germinal vecicle transplanting technology |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101045915A true CN101045915A (en) | 2007-10-03 |
Family
ID=38770837
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2006100709884A Pending CN101045915A (en) | 2006-03-29 | 2006-03-29 | Oocyte germinal vecicle transplanting technology |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101045915A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102766655A (en) * | 2012-06-20 | 2012-11-07 | 中国农业科学院北京畜牧兽医研究所 | Method for producing somatic cell cloned bovine blastocyst |
| US9938500B2 (en) | 2006-04-14 | 2018-04-10 | Astellas Institute For Regenerative Medicine | Hemangio-colony forming cells |
| US9988602B2 (en) | 2008-05-06 | 2018-06-05 | Astellas Institute For Regenerative Medicine | Methods for producing enucleated erythroid cells derived from pluripotent stem cells |
| CN111004775A (en) * | 2020-01-08 | 2020-04-14 | 中国医学科学院医学生物学研究所 | Method for physically removing oocyte zona pellucida |
| US10894065B2 (en) | 2012-12-21 | 2021-01-19 | Astellas Institute For Regenerative Medicine | Methods for production of platelets from pluripotent stem cells and compositions thereof |
| US12097223B2 (en) | 2011-11-30 | 2024-09-24 | Astellas Institute For Regenerative Medicine | Mesenchymal stromal cells and uses related thereto |
| US12209255B2 (en) | 2012-07-12 | 2025-01-28 | Astellas Institute For Regenerative Medicine | Mesenchymal-like stem cells derived from human embryonic stem cells, methods and uses thereof |
-
2006
- 2006-03-29 CN CNA2006100709884A patent/CN101045915A/en active Pending
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9938500B2 (en) | 2006-04-14 | 2018-04-10 | Astellas Institute For Regenerative Medicine | Hemangio-colony forming cells |
| US11566228B2 (en) | 2006-04-14 | 2023-01-31 | Astellas Institute For Regenerative Medicine | Hemangio-colony forming cells |
| US9988602B2 (en) | 2008-05-06 | 2018-06-05 | Astellas Institute For Regenerative Medicine | Methods for producing enucleated erythroid cells derived from pluripotent stem cells |
| US12097223B2 (en) | 2011-11-30 | 2024-09-24 | Astellas Institute For Regenerative Medicine | Mesenchymal stromal cells and uses related thereto |
| CN102766655A (en) * | 2012-06-20 | 2012-11-07 | 中国农业科学院北京畜牧兽医研究所 | Method for producing somatic cell cloned bovine blastocyst |
| US12209255B2 (en) | 2012-07-12 | 2025-01-28 | Astellas Institute For Regenerative Medicine | Mesenchymal-like stem cells derived from human embryonic stem cells, methods and uses thereof |
| US10894065B2 (en) | 2012-12-21 | 2021-01-19 | Astellas Institute For Regenerative Medicine | Methods for production of platelets from pluripotent stem cells and compositions thereof |
| US11400118B2 (en) | 2012-12-21 | 2022-08-02 | Astellas Institute For Regenerative Medicine | Methods for production of platelets from pluripotent stem cells and compositions thereof |
| US12076347B2 (en) | 2012-12-21 | 2024-09-03 | Astellas Institute For Regenerative Medicine | Methods for production of platelets from pluripotent stem cells and compositions thereof |
| US12109239B2 (en) | 2012-12-21 | 2024-10-08 | Astellas Institute For Regenerative Medicine | Methods for production of human hemogenic endothelial cells from pluripotent stem cells and compositions thereof |
| CN111004775A (en) * | 2020-01-08 | 2020-04-14 | 中国医学科学院医学生物学研究所 | Method for physically removing oocyte zona pellucida |
| CN111004775B (en) * | 2020-01-08 | 2023-05-02 | 中国医学科学院医学生物学研究所 | Method for physically removing oocyte zona pellucida |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101045915A (en) | Oocyte germinal vecicle transplanting technology | |
| CN101372682B (en) | Construction method of Epinephelus fuscoguttatus fin cell line | |
| CN202968563U (en) | Airlift biochemical culture device | |
| CN103805565B (en) | Hippocampal neuron is separated and primary culture method and reagent | |
| CN106367346A (en) | Mesenchymal stem cell extraction and perfusion culture system | |
| CN106867961A (en) | A kind of induced multi-potent stem cell forms mesoblastemic inducing culture and abductive approach | |
| CN201634683U (en) | Device for separating primary cells from tissue | |
| CN113637631B (en) | Extraction and culture method of rat peritoneal mesothelial cells | |
| CN104726339B (en) | A kind of immobilization cultural method of microalgae | |
| CN103305401A (en) | Stem cell extraction device | |
| CN104480153B (en) | The method for promoting micro- quasi- ball algae largely to accumulate high unsaturated fatty acid | |
| CN114921402A (en) | Method for extracting adult rat and mouse heart fibroblast | |
| CN208308829U (en) | A kind of electronic ovarian follicle revulseur | |
| CN207836420U (en) | A kind of sugarcane tissue culture device | |
| CN206736228U (en) | A kind of microbial cultivation device being easily installed | |
| CN202968544U (en) | Continuous fermentation propagation device for reducing contamination probability | |
| CN106978392A (en) | A kind of method of megalobrama amblycephala bone and its cells culture | |
| CN102140436A (en) | Culture solution for differentiating adipose-derived stromal cells into myocardial cells and preparation method thereof | |
| CN208023014U (en) | A kind of cell dissociation device | |
| CN214735822U (en) | Combined cell culture plate | |
| CN206599564U (en) | One kind is used for plant disease biological and ecological methods to prevent plant disease, pests, and erosion incubator | |
| CN107964531B (en) | Preparation method of duckweed living protoplast | |
| CN111471642B (en) | A method for efficiently promoting the proliferation of synovial cells based on tensile force | |
| CN211771343U (en) | Culture apparatus of gamma-aminobutyric acid lactic acid bacteria | |
| CN202465712U (en) | Stem cell isolated culturing device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |