CN201832181U

CN201832181U - Three-dimensional carrier

Info

Publication number: CN201832181U
Application number: CN2010202561258U
Authority: CN
Inventors: 吴世凯; 阳小卫; 姜舒; 郑意端; 胡祥
Original assignee: Shenzhen Beike Biotechnology Co Ltd
Current assignee: Shenzhen Beike Biotechnology Co Ltd
Priority date: 2010-07-09
Filing date: 2010-07-09
Publication date: 2011-05-18
Anticipated expiration: 2020-07-09

Abstract

The utility model provides an implantable coated three-dimensional carrier. The three-dimensional carrier comprises a carrier framework and a transparent film coated on the inner surface of the carrier framework. The transparent film cannot affect the free exchange of micromolecules, but can prevent the exchange of cells and high-molecular weight proteins, thus ensuring that the carrier cannot be degraded. The three-dimensional carrier has the advantages of simple structure, economy and applicability, and can effectively prevent immunoreaction and rejection which occur in the process of transplantation.

Description

A kind of three-dimensional carrier

[technical field]

This utility model bio-carrier technical field is specifically related to a kind of implantable toga three-dimensional carrier.

[background technology]

Along with the value of stem cell in clinical embodies, the transplantation immunity problem becomes the main difficulty of restriction stem cell clinical practice, how carrying out variant cell to transplant and not producing significant immunological rejection is a hot subject that presses for solution during present stem cell is used, in recent years, people adopt three-dimensional carrier to cultivate the method for transplanted cells, adopt sodium alginate, collagen, gelatin etc. are as the cell support material, peripheral parcel one deck poly-D-lysine or chitosan are as the bag quilt, cell both can be grown in the bag quilt, can be carried out the exchange and then the growing multiplication of small-molecule substance with the external world by bag again, this cultivates for cells in vitro good idea is provided, but in zoopery and clinical experiment, because poly-D-lysine or chitosan can be degraded by organism gradually, support material in the encapsulation still can directly contact with biological tissue with cell, thereby produce immunoreation or rejection, this just makes tissue repair be affected, therefore the patient may need the secondary reparation or cause slight illness more serious, brings huge financial burden for individual and society.

[utility model content]

In order to solve the deficiency of the above-mentioned technical problem that exists in the prior art, this utility model provides a kind of toga three-dimensional carrier that is difficult for being degraded that is used for tissue repair.

The technical scheme that the existing technical problem that solves this utility model adopts is: a kind of implantable toga three-dimensional carrier is provided, and described three-dimensional carrier comprises carrier framework and is coated in the semipermeable membrane of carrier framework inner surface.

Preferably: described semipermeable membrane is organic ether sulfone (PES) film, perhaps polysulfones (PSF) film, perhaps cellulose acetate (CA) film, perhaps polyvinylidene fluoride (PVDF) film, perhaps polypropylene (PP) film, perhaps mixed cellulose ester (CAN) film.

Of the present utility model further the improvement is that described carrier body is spherical, column or irregularly shaped; Be full of in the described carrier framework flowable bio-matrix and active substance are arranged.。

Of the present utility model further the improvement is that described carrier film thickness is 0.1--3mm.

The beneficial effects of the utility model are: 1, this utility model preparation process is simple, can finish under the normal temperature and pressure, is fit to large-scale production.

2, carrier film of the present utility model does not influence and micromolecularly freely exchanges, but can stop the exchange of cell and large molecular weight protein, has guaranteed that carrier is difficult for being degraded, and can effectively avoid the immunoreation and the rejection that produce in the migration process.

[description of drawings]

Fig. 1 is the sketch map of spherical sodium alginate carrier of the present utility model;

Fig. 2 is the sketch map of toga three-dimensional carrier of the present utility model.

[specific embodiment]

Below in conjunction with the description of drawings and the specific embodiment this utility model is further specified.

As shown in Figure 1 and Figure 2, this utility model provides a kind of implantable toga three-dimensional carrier, comprises carrier body 2 and carrier film 1, and carrier body 2 is wrapped in the carrier film 1; Carrier body 2 comprises bio-matrix 4 and active substance 3; Carrier body 2 is spherical, column or irregularly shaped; Carrier film 1 thickness is 0.1--3mm.

Active substance 3 is cells, or the activated protein factor, or the combination of the cell and the activated protein factor.Described cell comprises stem cell, CFU-GM and the one-tenth somatic cell thereof of various tissues, that is: islet cells, hematopoietic stem cell, mescenchymal stem cell, neural stem cell, endothelial progenitor cells, navel blood stem cell, Cord Blood Mononuclear Cell, retina cell; The described activated protein factor comprises interleukin, interferon, colony stimulating factor, somatomedin and chemotactic cytokine.Cell can be to have to be suitable for applying any kind of useful effectiveness biologically.Fetal tissue, the cambium of the cell that representative instance includes but not limited to break up fully, anchorage-dependent cell, birth tissue, not exclusively differentiation, do not rely on adherent transformant, cell line etc.More specifically but not exclusive ground, cell can comprise insulin production cell (for example islets of langerhans), adrenal pheochromocytoma, antibody secreting cell, fibroblast (especially utilizing genetic engineering to be treated to the fibroblast that produces the reorganization nerve growth factor), astrocyte and/or β cell line; Described bioactive substance can be any active factors, for example neurotransmitter, neuromodulator, catecholamine, somatomedin, cofactor, trophic factors and hormone, active substance also can be analog, agonist, derivant or have active active factors fragment that active substance can also be the inhibitor of bioactive substance.

The preparation material of bio-matrix 4 comprises collagen, sodium alginate, chitin, hyaluronic acid, gelatin, fibrin.

The preparation material of carrier film 1 is the macromolecular material with better biocompatibility, can be in polyether sulfone (PES), polysulfones (PSF), cellulose acetate (CA), polyvinylidene fluoride (PVDF), polypropylene (PP), mixed cellulose ester (CNA) and the polyurethane any.Non-polar material need be handled through polarity, and the preferred synthetic material of organizational project is the synthesized polymer body, comprises the oligomer, homopolymer and the copolymer that are come by sudden reaction and polycondensation reaction.Seal after with it parcel according to the carrier body shape, mouth-sealing method adopts heat-sealing mouthful or machinery to seal, and is stored in the culture fluid, needs wash with normal saline before using.When in this three-dimensional carrier implanting tissue, the carrier inner cell can the normal growth metabolism, but the suppressed by vector film intercepts directly contact tissue cell, greatly reduces the probability of generation immunologic rejection.

The preparation method of spherical sodium alginate carrier: with concentration is 1 * 10 ⁵～5 * 10 ⁶The MSC of/ml mixes with 10ml 2% sodium alginate, and mixture is cross-linked into microsphere by nozzle machine and CaCl2; Microsphere is transferred in the aseptic centrifuge tube of 50ml, adopts following sterile solution washing 0.55%CaCl successively ₂, 0.28%CaCl ₂, 0.85% normal saline, 0.1%CHES (2-N cyclic group amino-ethyl sulfonic acid), 1.1%CaCl ₂Next, and microsphere and 0.05% poly-D-lysine (MW 15000, Fluka, and USA) crosslinked 6min adopts 0.1%CHES, 1.1%CaCl then successively ₂, the washing of 0.85% normal saline, reuse 0.03% sodium alginate bag is formed layer structure by 4min; Microsphere after the washing is dissolved kernel 6min with the 0.55mmol/L sodium citrate; Use the serum-free medium washed twice, insert in the normal culture medium and cultivate; Preparing material with carrier film at last seals it parcel back with machinery.

Toga three-dimensional carrier preparation method two: with concentration is 1 * 10 ⁵～5 * 10 ⁶Direct and the host material mix homogeneously of/ml cell, the injection carrier film prepares material and seals, and mouth-sealing method can adopt heat-sealing mouth or machinery to seal.

Toga three-dimensional carrier preparation method three: collecting concentration is 1 * 10 ⁵～5 * 10 ⁶/ ml cell; Chitosan is dissolved in the acetic acid solution of 10ml/L, being prepared into concentration is the chitosan acetic acid solution of 2g/L, removes by filter impurity, and adding concentration is that the NaOH solution of 20mol/L is separated out chitosan, it is extremely neutral with the tri-distilled water washing to filter the back, obtains the purification chitosan through cold drying then.Carboxymethyl chitosan is dissolved in the normal saline, and with disposable pin type filter (0.22um) filtration sterilization; Utilize the high-voltage electrostatic field to become bag apparatus, the sodium alginate soln that with concentration is 17.5g/L is at voltage 3.0 * 10 ⁴V, fltting speed 50mm/h and liquid level are sprayed to 0.12mol/LCaCl under the 50mm condition ₂In the solution, form the calcium alginate gel microgranule; With microgel particle and 2g/L chitosan solution reaction 10min, make microgranule outer wrapping one deck chitosan; Add the 1.5g/L sodium alginate soln then, in and the microparticle surfaces excess charges; Obtain the sodium alginate-chitosan microsphere with 55mmol/L sodium citrate liquefaction capsule-core at last.Pack into this microsphere in the semipermeable membrane bag and seal.

Toga three-dimensional carrier preparation method four: get the good osteoblast of growth conditions and digest centrifugally, add in the gelatin solution (2.0% (w/v) is dissolved in the DMEM culture medium) and make cell resuspended, adjusting cell density is 2 * 10 ⁶Cells/mL.In incubator, hatch 30min, osteoblast is fully contacted with gelatin.In sodium alginate: the ratio of gelatin=3: 1 adds sodium alginate soln (2.0% (w/v)), behind the mix homogeneously, joins in the syringe, and this moment, cell density was 5 * 10cells/mL.Start the microcapsule preparation facilities,, under the flow velocity 40mL/h condition, drip the 1mL cell mixture to 30mLCaCl at voltage 6kV ₂Carry out gelation reaction in (1.1% (w/v)) solution, form and carry cell gelatin sodium alginate micro gel bead.Then the glue pearl is added to overlay film reaction 10min in the chitosan solution (0.5% (w/v)), forms and carry cell microcapsule.With PBS washing three times, the reuse serum-free medium washs once.This microsphere is packed in the semipermeable membrane bag, mouthful sew up with suture along the place.

Above content be in conjunction with concrete preferred implementation to further describing that this utility model is done, can not assert that concrete enforcement of the present utility model is confined to these explanations.For this utility model person of an ordinary skill in the technical field, under the prerequisite that does not break away from this utility model design, can also make some simple deduction or replace, all should be considered as belonging to protection domain of the present utility model.

Claims

1. three-dimensional carrier is characterized in that: described three-dimensional carrier comprises carrier framework and is coated in the semipermeable membrane of carrier framework inner surface.

2. three-dimensional carrier according to claim 1 is characterized in that: described semipermeable membrane is an organic ether sulfone film, perhaps polysulfone membrane, perhaps cellulose acetate membrane, perhaps PVDF membrane, perhaps polypropylene screen, perhaps mixed cellulose ester membrane.

3. three-dimensional carrier according to claim 1 is characterized in that: described carrier framework is spherical, column or irregularly shaped; Be full of in the described carrier framework flowable bio-matrix and active substance are arranged.

4. according to each described three-dimensional carrier of claim 1～3, it is characterized in that: described carrier film thickness is 0.1--3mm.