CN105062953A - Method for three-dimensional induction of transformation of mesenchymal stem cells into islet cells - Google Patents
Method for three-dimensional induction of transformation of mesenchymal stem cells into islet cells Download PDFInfo
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Abstract
The invention relates to a method for three-dimensional induction of transformation of mesenchymal stem cells into islet cells. The method comprises the steps of: acquiring mesenchymal stem cells, mixing the mesenchymal stem cells with PM polypeptide hydrogel and then performing induction culture so as to obtain islet cells. The method provided by the invention utilizes the PM polypeptide hydrogel with a three-dimensional network structure to provide an environment simulating cell growth in human body for the mesenchymal stem cells, provides sufficient growth space and nutrition requirements for the mesenchymal stem cells, and is in favor of enhancing the transformation rate of mesenchymal stem cells and the quality of islet cells.
Description
Technical field
The present invention relates to tissue engineering technique field, particularly a kind of three-dimensional inducing mesenchymal stem cell is converted into the method for islet cells.
Background technology
Diabetes be insulin secretion relatively or the defect such as definitely not enough or the insulin receptor sugar, fat and the protein metabolism disorder disease that cause.Clinically, I type and part ii patients with type Ⅰ DM many employings subcutaneous insulin injections are treated.And cell therapy is the ideal treatment strategy of diabetes especially insulin-dependent diabetes mellitus (IDDM), especially for the diabetic subject losing islet cell function, implanting the islet cells of excreting insulin or its surrogate to be optimal methods for the treatment of.At present, islet cell transplantation treatment diabetes achieve some curative effects, but donor's cells originates, the difficulty such as deficiency, serious immunological rejection significantly limit the application of this therapy.
Stem cell has extremely strong self-renewal capacity and multi-lineage potential, is the desirable target cell of gene therapy.Carrying out suitable genetic modification to stem cell, make it the cell becoming energy excreting insulin, is one of ideal strategy for the treatment of insulin-dependent diabetes mellitus (IDDM).And mescenchymal stem cell is a kind of cell with multi-lineage potential, it can directional induction be the various kinds of cell such as insulin-like cell, chondrocyte.And mesenchymal cell is present in human multiple tissue; especially the mescenchymal stem cell of umbilical cord, placenta, adipose tissue-derived; there are wide material sources, be easy to gather, without advantages such as ethics problems; mass-producing can be induced to differentiate into insulin-like cell, for clinical treatment diabetes provide new technical scheme.
Be the mode of the adherent induction of the many employings of Islet-like clusters at present by mesenchyma stem cell differentiation induction.Because mescenchymal stem cell has stronger adherent property, cell aggregation is hindered agglomerating in two-dimentional inducing culture system, therefore inductivity is general lower, and induces the problems such as the cytoactive of the rear insulin-like cell obtained is lower, viable cell ratio is low, islet cells is not mature enough.And two step method or three-step approach, inducible factor uses more, and induction duration is generally 14-21 days, and long, the residual factor of induction duration is of a great variety, cytoactive is low, Induction Transformation rate is low, adds the risk of clinical application.
Summary of the invention
Technical problem to be solved by this invention is, in two-dimentional inducing culture system, there is the defects such as cytoactive is low, Induction Transformation rate is low for mescenchymal stem cell in prior art, provide a kind of three-dimensional inducing mesenchymal stem cell of the transformation efficiency and cytoactive that can improve mescenchymal stem cell to be converted into the method for islet cells.
The technical solution adopted for the present invention to solve the technical problems is: provide a kind of three-dimensional inducing mesenchymal stem cell to be converted into the method for islet cells, said method comprising the steps of:
Cultivate in substratum after mescenchymal stem cell is mixed with PM polypeptide hydrogel, and in culturing process, add inductor induce.
Be converted in the method for islet cells at three-dimensional inducing mesenchymal stem cell provided by the invention, described PM polypeptide hydrogel is dissolved in deionized water by PM polypeptide and obtains.
Be converted in the method for islet cells at three-dimensional inducing mesenchymal stem cell provided by the invention, in described PM polypeptide hydrogel, the massfraction of PM polypeptide is 0.1-0.75%.
Be converted in the method for islet cells at three-dimensional inducing mesenchymal stem cell provided by the invention, before described PM polypeptide hydrogel mixes with mescenchymal stem cell, also comprise the step adopting basic culture solution or complete culture solution to infiltrate described PM polypeptide hydrogel.
Be converted in the method for islet cells at three-dimensional inducing mesenchymal stem cell provided by the invention, before described PM polypeptide hydrogel mixes with mescenchymal stem cell, also comprise the step adopting the resuspended mescenchymal stem cell of complete culture solution, and make the concentration of mescenchymal stem cell be 1 × 10
4individual/mL-1 × 10
6individual/mL.
Be converted in the method for islet cells at three-dimensional inducing mesenchymal stem cell provided by the invention, described substratum is IMDM nutrient solution containing EGF, bFGF, ginkgo extract and FCS or DMEM nutrient solution.
Be converted in the method for islet cells at three-dimensional inducing mesenchymal stem cell provided by the invention, in described substratum, the concentration of EGF is the massfraction that the concentration of 10-20ng/ml, bFGF is 10-20ng/ml, the concentration of ginkgo extract is 10-20ng/ml and FCS is 2-10%.
Be converted in the method for islet cells at three-dimensional inducing mesenchymal stem cell provided by the invention, described inductor comprises nicotinamide and HGF.
Be converted in the method for islet cells at three-dimensional inducing mesenchymal stem cell provided by the invention, the concentration of described nicotinamide is the concentration of 10-20ng/ml, HGF is 10-20ng/ml.
Be converted in the method for islet cells at three-dimensional inducing mesenchymal stem cell provided by the invention, add described inductor in culturing process and carry out in induction step, induction time is 4-7 days.
Implement the method that three-dimensional inducing mesenchymal stem cell provided by the invention is converted into islet cells, following beneficial effect can be reached: utilize the environment that the gel peptide with tridimensional network provides to simulate human inner cell and grow for mescenchymal stem cell, for mescenchymal stem cell provides sufficient growing space and nutritional needs, be conducive to the amplification of mescenchymal stem cell and improve the cytoactive of mescenchymal stem cell.
Embodiment
For mescenchymal stem cell in solution prior art is in two-dimensional culture system, mescenchymal stem cell expanding effect is not good, the not high defect of activity, innovative point of the present invention is to provide a kind of three-dimensional inducing mesenchymal stem cell to be converted into the method for islet cells, by adding the PM polypeptide hydrogel with tridimensional network in the medium, human inner cell's growing environment can be simulated, for mescenchymal stem cell provides sufficient growing space and nutritional needs, thus achieve the object improving mescenchymal stem cell activity and amplification times.
A kind of three-dimensional inducing mesenchymal stem cell provided by the invention is converted into the method for islet cells, comprises the following steps: obtain mescenchymal stem cell, with inductor carry out inducing culture after being mixed by mescenchymal stem cell with PM polypeptide hydrogel.
PM polypeptide hydrogel has tridimensional network, the microenvironment of human inner cell's growth can be simulated, increase the contact area of mescenchymal stem cell and substratum and oxygen, be conducive to the conduction of signal between cell, simultaneously, due to the attached cell that mescenchymal stem cell is stronger, the tridimensional network of PM polypeptide hydrogel, also for mescenchymal stem cell provides more sufficient apposition growth space, more be conducive to the amplification of mescenchymal stem cell, meanwhile, PM polypeptide hydrogel water-retentivity is comparatively strong, also can improve the activity of mescenchymal stem cell.
Preferably, PM polypeptide hydrogel of the present invention (also known as PuraMatrix gel peptide), purchased from BD company, a kind of peptide that PuraMatrix polypeptide is made up of 16 amino-acid residues, do not comprise obvious somatomedin or cytokine, self-assembly forms nanofibrous structures, in water-soluble solution, hydrophobic and ion amino acid combines the gel forming tridimensional network, i.e. PM polypeptide hydrogel, sufficient growing space can be provided for mescenchymal stem cell, expand the contact area of mescenchymal stem cell and nutrient solution, be conducive to the amplification of mescenchymal stem cell.At present, purposes disclosed in PM polypeptide hydrogel is for tissue regeneration, as bone filler, wound healing and for delivery system etc., and by after water-soluble for PM polypeptide hydrogel in the present invention, utilize its tridimensional network to carry out the inducing culture of mescenchymal stem cell, the culture effect of good mescenchymal stem cell can be obtained.
Particularly, the process of mescenchymal stem cell in dimensional culture system is:
1, with deionized water dissolving PM polypeptide, PM polypeptide hydrogel is obtained;
2, the PM polypeptide hydrogel of immersion step 1 acquisition;
3, obtain mescenchymal stem cell, and mix with the PM polypeptide hydrogel obtained in step 2;
4, cultivate in step 3 in substratum with the mixed mescenchymal stem cell of PM polypeptide hydrogel, and in culturing process, add inductor carry out inducing culture.
In step 1, because PM polypeptide is soluble in water, hydrophobic and ion amino acid combines and forms tridimensional network, and electrolyte substance can affect the solubleness of PM polypeptide in water, therefore, the deionized water removing electrolyte substance is adopted to carry out dissolved dilution PM polypeptide stoste in this step, preferably, in PM polypeptide hydrogel, the massfraction of PM polypeptide is 0.1%-0.75%, PM polypeptide hydrogel concentration is excessive, then the reticulated structure of PM polypeptide hydrogel formation is too fine and close, be unfavorable for the transduction of signal between the transmission of cytokine and cell, or cause the anoxic of cell, be unfavorable for the multiplication culture of cell.
In step 2, basic culture solution or complete culture solution can be adopted to carry out 2-4 time to the PM polypeptide hydrogel in step 1 infiltrate, be dissolved in PM polypeptide hydrogel to make the cytokine in nutrient solution and nutritional factor part, to increase the cellular affinity on PM polypeptide hydrogel surface, be conducive to cell attachment, and make the cytokine in basic medium and nutritional factor part be dissolved in PM polypeptide hydrogel, maintain Growth of Cells.
In step 3, because P3 mescenchymal stem cell propagation is more vigorous, cytoactive is comparatively strong, and preferentially choosing P3 in the present invention for mescenchymal stem cell, is 1 × 10 by complete culture solution re-suspended cell concentration
4-1 × 10
6individual/ml, adds in the PM polypeptide hydrogel solution in step 2 by cell suspension, leave standstill, treat that cell mixes with PM polypeptide hydrogel is full and uniform.
In step 4, the substratum adopted is DMEM nutrient solution or IMDM nutrient solution, EGF (Urogastron) wherein containing 10-20ng/mL, the bFGF (Prostatropin) of 10-20ng/mL, 10-20ug/mL ginkgo extract and 2-10%FCS (tire calf serum), after successive induction 4-7 days, add the inductor of HGF containing 5-20ng/mL nicotinamide/and 5-20ng/mL again, successive induction 4-7 days again, within every two days, change liquid 1 time, until mescenchymal stem cell is converted into islet cells.Be understandable that, the inductor adopted in this step is also not limited thereto, and the induction broth that mescenchymal stem cell can be converted into islet cells in prior art all uses the present invention.
Wherein, EGF can promote the propagation of mescenchymal stem cell, and the formation of islet cells group.BFGF can not only promote the propagation of mescenchymal stem cell and the increase of cell volume, also contributes to mescenchymal stem cell and changes into islet cells.Ginkgo extract extracts according to international standard extraction process, wherein containing abundant antioxidase, flavonoid and bilobalide etc., the oxyradical accumulated in cell cultivation process can be neutralized, cell fission can be made like this to arrive very high density and not dead, thus be conducive to the propagation of mescenchymal stem cell, reduce the oxygenizement of cell, improve the level of glutathion inside cell, scavenging activated oxygen, prevent the apoptosis that cell is too early in the process of inducing or increase, be conducive to the conversion of mescenchymal stem cell; Also can affect the metabolism of zooblast cholesterol.FCS contains the necessary nutrition of abundant Growth of Cells; hormone to maintaining cell index growth is provided; be cell attachment, spread over required Factor Source on culture medium; and proteinase inhibitor is provided; make to remain trypsin inactivation when passage; Cell protection preserves from, and meanwhile, plays the effect of potential of hydrogen damping fluid.
And the Main Function of nicotinamide participates in respiratory chain, participate in respiratory and the electron transfer process of cell proliferation conversion, promote that the propagation of cell transforms, nicotinamide is a part of composition NAD+ and NADP+, NAD+ and NADP+ of composition, by electron transport chain, transmits H+, synthesis ATP; Nicotinamide can regulate the ratio of intracellular ATP/ADP simultaneously, thus cause the closedown of ATP sensitive potassium channel on cytolemma, film generation depolarize, then cause voltage-dependent ca channel open, flow of calcium ions, triggers islet β cell Regular Insulin, can promote that mesenchyma stem cell to pancreatic islet like cell breaks up, meanwhile, islet cell aysmature syndrome is promoted.
HGF is in the process of insulin-like cell in mescenchymal stem cell cytodifferentiation; to the amplification important role of cell while playing a protective role for cell; growth and the differentiation of islet cells can be promoted, improve activity and the ripening degree of islet cells further.
Therefore, in dimensional culture system, the method for islet cells is converted into by mescenchymal stem cell provided by the invention, the transformation efficiency of mescenchymal stem cell can not only be improved, improve cytoactive, and the islet cell mass that the present invention obtains is more, and activity is comparatively strong, ripening degree is better, and islet cells quality is stronger.
In order to make object of the present invention, technical scheme and advantage clearly understand, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
Embodiment 1
Mescenchymal stem cell provided by the invention is converted into the method for islet cells in dimensional culture system, comprises the following steps:
1, preparation is the PM polypeptide hydrogel of 0.5%PM polypeptide containing massfraction;
2, the PM polypeptide hydrogel solution of immersion step 1 acquisition;
3, obtain P3 for mescenchymal stem cell, mix with the PM polypeptide hydrogel in step 2;
4, cultivate in step 3 in substratum for mescenchymal stem cell with the mixed P3 of PM polypeptide hydrogel, and add inductor and carry out inducing culture.
In step 1, PM polypeptide is put into ultrasound bath pot sonic oscillation 30min, wherein, the water that this water-bath adopts is deionized water; If produce bubble in oscillatory process, be transferred to centrifuge tube centrifugal (4000 turns/5min) and remove.According to the final concentration of required PM polypeptide, get EP pipe (the eppendorf pipe of 1.5mL, indicate the centrifuge tube of respective concentration), with the PM polypeptide hydrogel of the PM polypeptide stoste preparation desired concn of 25ul deionized water dilution 25ul, preparation PM peptide masses mark is the PM polypeptide hydrogel of 0.5%.
It is uneven that existence due to bubble makes PM polypeptide hydrogel disperse in deionized water, affect the amplification density of mescenchymal stem cell, and bubble structure is unstable, can break at any time, easy destruction is attached to the mescenchymal stem cell on PM polypeptide hydrogel, therefore, in this process, avoid producing bubble as far as possible.
In step 2, the 0.5%PM polypeptide hydrogel obtained in step 1 is inoculated in cell culture container, as in culture plate, the disposable PM polypeptide hydrogel 50 μ L injecting respective concentration from culture hole center fast, avoid producing bubble as far as possible, if any bubble, puncture by fine needle, and ensure that high density PM polypeptide hydrogel is paved with at the bottom of hole, culture plate is placed in 37 DEG C of incubators and leaves standstill 15min.
Then, inject 200 μ L basic culture solutions along cultivation hole wall lentamente with 200 μ L liquid-transfering guns are careful, avoid producing bubble; Be placed in 37 DEG C of incubators standing promotion PM polypeptide hydrogel to solidify; Carefully inhale with liquid-transfering gun after 60min and abandon 150 μ L basic culture solutions, add the basic culture solution that 150 μ L are new, repeat this operation 2 times, every minor tick 60min, PM polypeptide hydrogel is infiltrated, is dissolved in PM polypeptide hydrogel to make the cytokine in basic medium and nutritional factor part.
In step 3, acquisition P3 for the process of mescenchymal stem cell is:
1) people's umbilical cord is obtained;
Take fully moon umbilical cord of Cesarean esction fetus about 4 ~ 6cm long, aseptically pump Cord blood, be placed in PBS damping fluid 4 DEG C preservation, process in 4h.
2) separation and purification mescenchymal stem cell
Long for about 4 ~ 6cm umbilical cord is put into super clean bench, and remove arteriovenous and adventitia, the PBS damping fluid of PH=7.4 carries out first rinsing 2-3 time, is cut into 1cm segment, then with PBS damping fluid repeatedly rinsing to remove blood as far as possible, be finally cut into meat gruel shape.
The umbilical cord of meat gruel shape is moved into 50mL centrifuge tube, add 5mL through 37 DEG C of preheatings, massfraction be 0.1% collagenase II, put into 37 DEG C of shaking baths vibration digestion 180min.Wherein, collagenase II is obtained by the PBS damping fluid of 0.1g collagenase digestion in 100ml.
Then, then add the DMEM/F12 of 30mL, repeatedly blow and beat, the sieved filter of cell, collect filtrate; Filtrate, with the centrifugal 10min of 1000 turns/min, is abandoned supernatant, is namely obtained mescenchymal stem cell.
3) Secondary Culture
By step 2) in the mescenchymal stem cell that is centrifugation down by 1 × 10
6the density of individual/mL is inoculated in culturing bottle with the DMEM/F12 containing 10%FBS (foetal calf serum), moves in incubator and cultivates.Next day, half amount changed liquid first, and every 3d changes liquid 1 time later.
After cell reaches fusion, prepare with trypsin trypsin solution: PBS damping fluid 200mL, trypsinase 0.5g, EDTA (ethylenediamine tetraacetic acid (EDTA)) 0.04g) incomplete digestion carries out Secondary Culture.
4) step 3 is got) the middle P3 obtained is for mescenchymal stem cell, and after tryptic digestion, resuspended adjustment cell density is 1 × 10
5individual/ml, adds in the 0.5%PM polypeptide hydrogel in step 2 by cell suspension, leave standstill 10 minutes, treat that cell mixes with PM polypeptide hydrogel is full and uniform.
In step 4, add the EGF of 5ml containing l0ng/mL in the mixture of the mescenchymal stem cell in step 3 and PM polypeptide hydrogel, the IDMEM nutrient solution of the bFGF of l0ng/mL, 10ug/mL ginkgo extract and 2%FCS, is placed in 5%CO by culture plate
2, in 37 DEG C of incubators, continue cultivation 4 days, within the 4th day, add the inductor of the HGF containing 10ng/mL nicotinamide and 10ng/mL, then successive induction 4 days, within every two days, change liquid 1 time, until mescenchymal stem cell is converted into islet cells.
For verifying that the method that mescenchymal stem cell provided by the invention is converted into islet cells in three-dimensional inducing culture system has outstanding significance effect further, further describe below by way of specific experiment.
Experimental group: the cell that the embodiment of the present invention 1 obtains;
Control group: with the embodiment of the present invention 1 dimensional culture system unlike, this group by 2D culture system is obtained cell.
Testing process:
Respectively following operation is performed to experimental group and control group: centrifugal collecting cell (5-10 × 10
6individual/ml, 0.5ml) wash twice with PBS damping fluid after add sour ethanolic soln 1ml (preparing 10% Glacial acetic acid with dehydrated alcohol), 4 DEG C of extractings are spent the night, by cell Ultrasonic Cell Disruptor crash cells, surpass five times, each 10s, power is 60-100W, the centrifugal 10min of 4000r/min at 4 DEG C, collects supernatant and is stored in-70 DEG C of refrigerators.Insulin level in supernatant and endochylema supernatant is measured with Regular Insulin radioimmunological kit agent box.
Table 1
| Detected object | Insulin level (μ IU/ml) in endochylema | Insulin level (μ IU/ml) in supernatant |
| Control group | 1800.33±209.07 | 4800.56±900.27 |
| Experimental group | 3712.77±312.86 | 7328.37±1212.63 |
Detected result: as shown in table 1, the insulin level in experimental group endochylema and supernatant is all higher than control group, and illustrate thus, the islet cells quality obtained by the present invention is higher, can produce more Regular Insulin.
In sum, mescenchymal stem cell provided by the invention is converted into the method for islet cells in dimensional culture system, not only simple to operation, and adopt the PM polypeptide hydrogel with tridimensional network, for cell provides a three dimensional growth space, can the growing environment of analog cell in human body, for cell provides sufficient growing space, and increase the contact area of mescenchymal stem cell and nutrient solution and oxygen and intercellular conduction, can not only promote that mescenchymal stem cell is converted into islet cells, and contribute to the quality improving islet cells.
Above embodiments of the invention are described; but the present invention is not limited to above-mentioned embodiment; above-mentioned embodiment is only schematic; instead of it is restrictive; those of ordinary skill in the art is under enlightenment of the present invention; do not departing under the ambit that present inventive concept and claim protect, also can make a lot of form, these all belong within protection of the present invention.
Claims (10)
1. three-dimensional inducing mesenchymal stem cell is converted into a method for islet cells, it is characterized in that: said method comprising the steps of:
Cultivate in substratum after mescenchymal stem cell is mixed with PM polypeptide hydrogel, and in culturing process, add inductor induce.
2. three-dimensional inducing mesenchymal stem cell according to claim 1 is converted into the method for islet cells, it is characterized in that, described PM polypeptide hydrogel is dissolved in deionized water by PM polypeptide and obtains.
3. three-dimensional inducing mesenchymal stem cell according to claim 2 is converted into the method for islet cells, it is characterized in that, in described PM polypeptide hydrogel, the massfraction of PM polypeptide is 0.1-0.75%.
4. three-dimensional inducing mesenchymal stem cell according to claim 1 is converted into the method for islet cells, it is characterized in that, before described PM polypeptide hydrogel mixes with mescenchymal stem cell, also comprise the step adopting basic culture solution or complete culture solution to infiltrate described PM polypeptide hydrogel.
5. three-dimensional inducing mesenchymal stem cell according to claim 1 is converted into the method for islet cells, it is characterized in that, before described PM polypeptide hydrogel mixes with mescenchymal stem cell, also comprise the step adopting the resuspended mescenchymal stem cell of complete culture solution, and make the concentration of mescenchymal stem cell be 1 × 10
4individual/mL-1 × 10
6individual/mL.
6. three-dimensional inducing mesenchymal stem cell according to claim 1 is converted into the method for islet cells, it is characterized in that, described substratum is IMDM nutrient solution containing EGF, bFGF, ginkgo extract and FCS or DMEM nutrient solution.
7. three-dimensional inducing mesenchymal stem cell according to claim 6 is converted into the method for islet cells, it is characterized in that, in described substratum, the concentration of EGF is the massfraction that the concentration of 10-20ng/ml, bFGF is 10-20ng/ml, the concentration of ginkgo extract is 10-20ng/ml and FCS is 2-10%.
8. three-dimensional inducing mesenchymal stem cell according to claim 1 is converted into the method for islet cells, it is characterized in that, described inductor comprises nicotinamide and HGF.
9. three-dimensional inducing mesenchymal stem cell according to claim 8 is converted into the method for islet cells, it is characterized in that, the concentration of described nicotinamide is the concentration of 10-20ng/ml, HGF is 10-20ng/ml.
10. three-dimensional inducing mesenchymal stem cell according to claim 1 is converted into the method for islet cells, it is characterized in that, add described inductor in culturing process and carry out in induction step, induction time is 4-7 days.
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| CN105477017A (en) * | 2015-12-03 | 2016-04-13 | 深圳爱生再生医学科技有限公司 | Mixed stem cell preparation for treating diabetic foot and preparation method of mixed stem cell preparation |
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| CN113174408A (en) * | 2021-04-28 | 2021-07-27 | 吉林大学 | Islet cells differentiated from stem cells, method, compound and application |
| CN113181217A (en) * | 2021-04-28 | 2021-07-30 | 吉林大学 | Cell scaffold, islet cell carrying compound thereof, preparation method and application |
| CN114045253A (en) * | 2021-10-28 | 2022-02-15 | 东南大学 | Stem cell and islet beta cell co-culture method based on composite hydrogel |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105477017A (en) * | 2015-12-03 | 2016-04-13 | 深圳爱生再生医学科技有限公司 | Mixed stem cell preparation for treating diabetic foot and preparation method of mixed stem cell preparation |
| WO2019093468A1 (en) * | 2017-11-10 | 2019-05-16 | 富士フイルム株式会社 | Method for producing insulin-producing cells from mesenchymal stem cells, insulin-producing cells, cell structure, and pharmaceutical composition |
| CN111344393A (en) * | 2017-11-10 | 2020-06-26 | 富士胶片株式会社 | Method for producing insulin-producing cell from mesenchymal stem cell, insulin-producing cell, cell construct, and pharmaceutical composition |
| JPWO2019093468A1 (en) * | 2017-11-10 | 2021-01-14 | 富士フイルム株式会社 | Methods for Producing Insulin-producing Cells from Mesenchymal Stem Cells, Insulin-producing Cells, Cell Structures and Pharmaceutical Compositions |
| JP2022079721A (en) * | 2017-11-10 | 2022-05-26 | 富士フイルム株式会社 | Method for producing insulin-producing cells from mesenchymal stem cells, insulin-producing cells, cell structure, and pharmaceutical composition |
| JP7078939B2 (en) | 2017-11-10 | 2022-06-01 | 富士フイルム株式会社 | Methods for Producing Insulin-Producing Cells from Mesenchymal Stem Cells, Insulin-Producing Cells, Cell Structures and Pharmaceutical Compositions |
| JP7360672B2 (en) | 2017-11-10 | 2023-10-13 | 富士フイルム株式会社 | Method for producing insulin-producing cells from mesenchymal stem cells, insulin-producing cells, cell structures, and pharmaceutical compositions |
| CN113174408A (en) * | 2021-04-28 | 2021-07-27 | 吉林大学 | Islet cells differentiated from stem cells, method, compound and application |
| CN113181217A (en) * | 2021-04-28 | 2021-07-30 | 吉林大学 | Cell scaffold, islet cell carrying compound thereof, preparation method and application |
| CN114045253A (en) * | 2021-10-28 | 2022-02-15 | 东南大学 | Stem cell and islet beta cell co-culture method based on composite hydrogel |
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