CN115896009A - Efficient low-cost culture medium for in-vitro differentiation of adipose-derived mesenchymal stem cells - Google Patents
Efficient low-cost culture medium for in-vitro differentiation of adipose-derived mesenchymal stem cells Download PDFInfo
- Publication number
- CN115896009A CN115896009A CN202211286114.8A CN202211286114A CN115896009A CN 115896009 A CN115896009 A CN 115896009A CN 202211286114 A CN202211286114 A CN 202211286114A CN 115896009 A CN115896009 A CN 115896009A
- Authority
- CN
- China
- Prior art keywords
- differentiation
- medium
- mesenchymal stem
- derived mesenchymal
- adipose
- 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
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides an improved differentiation medium which is used for in vitro differentiation of pig fat mesenchymal stem cells, has high efficiency and low cost and is safe for food. The invention adopts the improved differentiation medium to induce the adipose-derived mesenchymal stem cells to be differentiated into fat, and compared with the traditional differentiation medium, the invention can improve the relative expression quantity of PPAR gamma genes and proteins on the 5 th day of differentiation; increasing relative expression levels of FABP4 and Plun 1 genes and proteins at day 10 of differentiation; and the oil red O staining result proves that more and more lipid droplets are accumulated by using the improved differentiation culture medium to induce the adipogenic differentiation of the adipose-derived mesenchymal stem cells. In conclusion, the improved differentiation medium provided by the invention improves the adipogenic differentiation efficiency of the adipose-derived mesenchymal stem cells, and provides a more efficient, healthy and low-cost adipogenic differentiation medium for large-scale production of cultured meat.
Description
Technical Field
The invention belongs to the technical field of stem cell and animal cell culture meat, and particularly relates to a culture medium which is used for adipose mesenchymal stem cells, has higher efficiency of in-vitro differentiation and lower cost price, and can be safely used for food.
Background
The cell culture meat is obtained by culturing stem cells in vitro according to the growth mechanism of animal muscle and fat, and the cell culture meat is directly used for industrially producing meat without animal culture. The cell culture meat as a subversive meat production mode provides a new way for supplementing protein supply of future meat and realizing green production of meat. According to the measurement, compared with the traditional animal husbandry, the cell culture meat industry can reduce the energy consumption by 35 to 60 percent, occupy less land by 98 percent and produce less greenhouse gases by more than 80 percent.
Adipose-derived mesenchymal stem cells (Adipose-derived mesenchymal stem cells) are stem cells having the ability to proliferate and differentiate into mature adipocytes. In recent years, various induction methods and induction factors have succeeded in inducing and differentiating adipose-derived mesenchymal stem cells, and most of the induction methods and induction factors adopt combined induction of hormones such as insulin, 3-isobutyl-1-methylxanthine, dexamethasone, rosiglitazone and indomethacin.
Similarly, in the in vitro production process of the cultured meat, a method for inducing adipose-derived mesenchymal stem cells to efficiently differentiate into mature adipocytes needs to be established, so that the selection of an excellent cell differentiation medium is a key part in the production process of the cultured meat, and the generation of lipid droplets is directly influenced.
The culture medium formula for inducing the differentiation of the adipose-derived mesenchymal stem cells into mature adipocytes is widely used at present, and is prepared by adding 10% bovine serum and 1% double antibody into a DMEM/F12 basal medium and performing combined induction by using five inducers, namely 3-isobutyl-1-methylxanthine, insulin, dexamethasone, indomethacin and rosiglitazone. The culture medium can achieve a better differentiation effect about 10 days of differentiation. However, the culture medium has the problems of strong potential toxicity, non-compliance with food safety related requirements, high cost and the like due to the addition of chemical components such as 3-isobutyl-1-methylxanthine, dexamethasone, indomethacin and rosiglitazone. In addition, the differentiation efficiency of adipocytes under this medium condition was not satisfactory. Therefore, it is important to develop a culture medium which can be safely used for food, reduce the price cost and promote the efficient differentiation of adipose-derived mesenchymal stem cells by using oleic acid, common vitamin C, vitamin B, vitamin K and low-concentration insulin.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to replace four components of 3-isobutyl-1-methylxanthine, dexamethasone, indomethacin and rosiglitazone in the traditional differentiation medium and reduce the using amount of insulin in the traditional differentiation medium, and provides a set of improved differentiation medium which can be safely used for food, reduce the price cost and promote the efficient differentiation of adipose mesenchymal stem cells and an application method thereof through oil oleic acid, vitamin C, vitamin B, vitamin K and low-concentration insulin.
The invention provides an improved differentiation medium which can be safely used for food, reduces the price cost and promotes the efficient differentiation of adipose-derived mesenchymal stem cells. The improved differentiation culture medium added with the cell culture supplementary factors can be prepared by directly adding the cell culture supplementary factors into a general culture medium. The general culture medium can be selected from various conventional cell culture media, such as DMEM, MEM, DMEM/F12, F10 and the like, and the DMEM/F12 medium is used for adding cell culture supplementary factors in the embodiment of the invention.
The additional lipotropic differentiation factors may be selected from a plurality of the following cell-complementing factors: insulin, oil (soybean oil), vitamin C, vitamin B, vitamin K, oleic acid, etc.
Furthermore, the cell culture media added with the cell culture complementary factors all contain penicillin-streptomycin double-resistant solution, the penicillin content is 10000U/ml, and the streptomycin content is 10mg/ml.
In the cell culture medium added with the cell culture supplementary factors, the total concentration of the added cell culture supplementary factors is not less than 50mg/mL, and preferably 100-1500mg/mL.
Furthermore, in the cell culture medium added with the cell culture supplementary factors, the concentration of any added cell culture supplementary factors is in the range of 0.5ng/mL-0.2 mg/mL.
In a third aspect of the present invention, there is provided an application method of the aforementioned high-efficiency and low-cost adipogenic differentiation medium in the differentiation culture of adipose-derived mesenchymal stem cells in vitro, comprising the following steps:
1) And taking the purified adipose-derived mesenchymal stem cells.
2) Adding a recombinant human Fibroblast Growth Factor (bFGF) into a culture dish to amplify the adipose-derived mesenchymal stem cells to a density of more than 90%;
3) After absorbing the adipose-derived mesenchymal stem cell proliferation culture medium, replenishing a new cell culture medium added with adipogenic differentiation factors again, and continuing to culture;
4) After 8 days, absorbing the culture medium, adding a differentiation maintaining culture medium only containing insulin, and continuing to culture;
5) After 10 days the adipocytes produced large lipid droplets.
Further, the environmental conditions for the cell culture are CO 2 Culturing at 37 deg.C in incubator with CO 2 CO in the incubator 2 The concentration was 5% (v/v).
Further, the adipose-derived mesenchymal stem cell proliferation medium used in step 2) was 10vol% of fetal bovine serum, 89vol% of F12 medium, 1vol% of penicillin streptomycin diabody and 5ng/ml of bFGF.
The existing adipose-derived mesenchymal stem cell differentiation medium contains a lot of chemical components which do not meet the food safety related requirements, has strong potential toxicity, is limited in the application of biochemistry, medicine, food science and the like, has low efficiency of inducing adipose-derived mesenchymal stem cell differentiation, and cannot fully utilize the differentiation potential of adipose-derived mesenchymal stem cells. Compared with the old formula, the adipose-derived mesenchymal stem cell differentiation medium added with the oleic acid and the vitamins can improve the relative expression of the PPAR gamma gene by 1.49 (+ -0.83) times on the 5 th day of differentiation and improve the relative expression of the FABP4 gene by 2.48 (+ -0.75) times on the 10 th day of differentiation. The relative expression of the Plin1 gene was increased 1.62 (. + -. 0.62) fold on day 10 of differentiation.
The relative expression level of PPAR gamma protein can be increased by 1.49 (+ -0.83) times on the 5 th day of differentiation, and the relative expression level of FABP4 protein can be increased by 1.97 (+ -0.06) times on the 10 th day of differentiation; the relative expression of the Plun 1 protein can be increased by 1.82 (+ -0.06) times at day 10 of differentiation. The cell differentiation effect at the terminal differentiation stage is better, more lipid droplets are formed by induced differentiation and are fuller, the cell differentiation culture medium can improve the cell differentiation percentage, the differentiation efficiency of the traditional differentiation culture medium on the tenth day of differentiation only reaches 11.67 percent (+ -0.02), the differentiation efficiency of the improved differentiation culture medium on the tenth day of differentiation can reach 29.38 percent (+ -0.02), and the very significant difference (P < 0.001) exists, and is 2.5 times of the differentiation efficiency of the traditional differentiation culture medium. The improvement of the differentiation performance provides a method for efficiently differentiating the adipose tissue-derived mesenchymal stem cells into mature adipose cells, reducing the compound without food safety use history, reducing the price cost and promoting the efficient differentiation of the adipose tissue-derived mesenchymal stem cells for 3D culture of adipose tissue-derived mesenchymal stem cell production cell culture meat.
The technical scheme of the invention has the following beneficial effects:
the key point of the production of the cell culture meat is that the adipose-derived mesenchymal stem cells enter a differentiation process, and the cells generate triglyceride and are condensed into lipid droplets, but the existing lipid differentiation promoting culture medium has the defects of no compound with a food safety use history, strong potential toxicity, low differentiation efficiency, high cost and the like, which means that the cell culture meat cannot be produced efficiently, stably and inexpensively on a large scale, and the industrialization process of the cell culture meat is seriously hindered.
The culture medium can be safely used for food, is low in price and cost, and can promote the efficient differentiation of the adipose-derived mesenchymal stem cells, and the addition of the oleic oil and the vitamin C, the vitamin B and the vitamin K replaces the use of the traditional inducer in the differentiation culture medium, so that the differentiation efficiency of the adipose-derived mesenchymal stem cells is improved in the process of inducing the differentiation of the adipose-derived mesenchymal stem cells in vitro, the differentiation potential of the adipose-derived mesenchymal stem cells is stimulated, and simultaneously, more triglyceride can be generated to form more lipid droplets. The adipose-derived mesenchymal stem cell differentiation medium with safe chemical components can also provide higher differentiation efficiency when 3D production of cell culture meat is carried out, more adipogenic differentiation related proteins are expressed, and cell culture meat with higher quality is produced.
Drawings
Fig. 1 is a bright field picture of a conventional differentiation medium and an improved differentiation medium inducing differentiation of adipose-derived mesenchymal stem cells to day 10. A is a bright field picture of the traditional differentiation medium induced differentiation to day 10. And B is a bright field picture of the improved differentiation medium induced differentiation to the 10 th day.
Fig. 2 shows the PPAR γ gene expression of differentiated adipose-derived mesenchymal stem cells on day 5 of differentiation induced by the conventional differentiation medium and the modified differentiation medium, respectively.
FIG. 3 shows that the expression of FABP4 gene in differentiated adipose-derived mesenchymal stem cells is induced by the conventional differentiation medium and the improved differentiation medium respectively at the 10 th day of differentiation.
FIG. 4 shows the expression of Plin1 gene in differentiated adipose-derived mesenchymal stem cells induced by the conventional differentiation medium and the improved differentiation medium respectively on day 10 of differentiation.
FIG. 5 shows that the conventional differentiation medium and the modified differentiation medium respectively induce the expression of PPAR γ proteins of differentiated adipose-derived mesenchymal stem cells on the 5 th day of differentiation.
FIG. 6 shows the expression of FABP4 and Plun 1 proteins of differentiated adipose-derived mesenchymal stem cells respectively at day 10 of differentiation induced by the conventional differentiation medium and the modified differentiation medium.
FIG. 7 is a comparison of cell differentiation percentages of a conventional differentiation medium and an improved differentiation medium, which induce differentiated adipose-derived mesenchymal stem cells, respectively, at day 10.
Detailed Description
The invention discovers that the improved differentiation medium can improve the relative expression of PPAR gamma gene by 1.49 (+ -0.83) times on the 5 th day of differentiation and improve the relative expression of FABP4 gene by 2.48 (+ -0.75) times on the 10 th day of differentiation compared with the traditional differentiation medium. The relative expression of the Plin1 gene was increased 1.62 (. + -. 0.62) fold on day 10 of differentiation.
The relative expression level of PPAR gamma protein can be increased by 1.49 (+ -0.83) times on the 5 th day of differentiation, and the relative expression level of FABP4 protein can be increased by 1.97 (+ -0.06) times on the 10 th day of differentiation; the relative expression of the Plun 1 protein can be increased by 1.82 (+ -0.06) times at day 10 of differentiation. The cell differentiation effect is better in the terminal differentiation stage, more and fuller lipid droplets are formed by induced differentiation, the cell differentiation medium improves the cell differentiation percentage, more cells generate new lipid droplets, the differentiation efficiency of the traditional differentiation medium reaches 11.67 percent (plus or minus 0.02) on the tenth differentiation day, and the differentiation efficiency of the improved differentiation medium reaches 29.38 percent (plus or minus 0.02) on the tenth differentiation day, which is 2.5 times that of the traditional differentiation medium. The improvement of the differentiation performance provides a more efficient and low-cost method for differentiating the adipose-derived mesenchymal stem cells into mature adipose cells more efficiently and producing cell culture meat by 3D culture of the adipose-derived mesenchymal stem cells.
The novel efficient and low-cost cell differentiation culture medium provided by the invention is a cell culture medium added with cell culture supplementary factors, and is consistent with the in-vitro culture method of normal adipose mesenchymal stem cells in other aspects.
All cell differentiation media (including positive controls) used in the following examples were DMEM/F12 basal media supplemented with 10% horse serum and 1% diabody cell culture media, and with small molecule substances (table 1), the modified differentiation media comprising components a, B, C, D, E, J; traditional differentiation media comprise components F, G, H, I, J.
The culture medium used in the proliferation phase in the following examples was DMEM/F12 basal medium supplemented with 10% fetal bovine serum and 1% diabody supplemented with 5ng/ml bFGF.
The cells used in the following examples were piglet adipose-derived mesenchymal stem cells, and further adherent cells.
The culture conditions used in the following examples were all culture at 37 ℃ in a CO2 incubator, and the CO2 concentrations were all 5% (v/v).
The detection methods employed in the following examples are, unless otherwise indicated, experimental methods, detection methods and preparation methods disclosed in the art.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
TABLE 1
Example 1 induced differentiation of porcine adipose-derived mesenchymal stem cells:
the experiment is divided into two groups, namely an existing adipogenic differentiation medium treatment group (positive control) added with a traditional inducer and an improved adipogenic differentiation medium treatment group provided by the invention, and the specific treatment method comprises the following steps:
1) Cell inoculation: pig adipose-derived mesenchymal stem cells before the P5 generation are inoculated into a 3.5cm culture dish according to the density of 60000-120000 cells/plate, and cultured by using a proliferation medium of DMEM/F12 containing 10% fetal bovine serum and added with 5ng/ml bFGF, and the culture medium is changed every two days.
2) Inducing differentiation: after the cells are proliferated to be converged, sucking out a proliferation culture medium, respectively cleaning the cells once by using a traditional differentiation culture medium and the improved differentiation culture medium, adding a DMEM/F12 culture medium containing 10% fetal calf serum into a negative control group, adding the traditional differentiation culture medium into a positive control group, adding the improved differentiation culture medium into an experimental group, carrying out induced differentiation on the cells, carrying out full liquid change once every 2 days, and observing the differentiation effect of differentiating to form lipid droplets every day.
Specifically, the differentiation medium used in the experimental group on days 1 to 7 is an improved differentiation medium for induced differentiation, and the DMEM/F12 medium containing 10% fetal calf serum for induced differentiation on days 8 to 10; the positive control group was induced to differentiate on days 1 to 4 using a conventional differentiation medium, on days 5 to 7 using a maintenance differentiation medium containing insulin only, and on days 8 to 10 using a DMEM/F12 medium containing 10% fetal bovine serum. Differentiation of negative control group on days 1 to 10 differentiation was induced uniformly with DMEM/F12 medium containing 10% fetal bovine serum.
3) The results show that: the improved differentiation medium can enhance the differentiation capacity of adipose-derived mesenchymal stem cells, improve the differentiation efficiency, and produce more and fuller lipid droplets (figure 1).
Example 2 oil red staining induced by differentiating porcine adipose-derived mesenchymal stem cells
1) Collecting a sample: mature adipocytes induced to differentiate by example 1 were aspirated from the medium, washed 1 time with PBS, and fixed with 4% paraformaldehyde overnight at 4 ℃.
2) Oil red dyeing: 4% paraformaldehyde was aspirated, washed 3 times with PBS for 5 minutes each, and washed in 60% isopropanol for 5 minutes. The sections were stained in oil red stain (prepared in isopropanol) for 15 minutes in the dark to make the lipid droplets appear red and soaked in 60% isopropanol for 1 minute.
Preparing oil red O working solution: diluting the stock solution 5% oil red O dye solution with triple distilled water according to the ratio of 3.
60% isopropyl alcohol: 30ml of isopropyl alcohol +20ml of triple distilled water, and stored at 4 ℃.
3) Cell differentiation rate statistics: oil red quantification was done using the software Image pro plus to analyze the lipid droplet area ratio in oil red staining, i.e. the percentage of the total area of red fat droplets over the whole 10 x Image area used (figure 8).
4) The results show that: the improved differentiation medium increases the percentage of cells differentiated, and allows more cells to generate new lipid droplets. The differentiation efficiency of the traditional differentiation medium on the tenth differentiation day is 11.67% (+ -0.02), and the differentiation efficiency of the improved differentiation medium on the tenth differentiation day is 29.38% (+ -0.02), which is 2.5 times that of the traditional differentiation medium.
Example 3 detection of Gene and protein level in differentiation induced by porcine adipose-derived mesenchymal Stem cells
1) And (3) gene level detection:
samples were taken on days 5 and 10 of the induced differentiation in step 3) according to the treatment method of example 1, and the PPAR γ gene expression level on day 5 and the FABP4 and the PLIN1 gene expression level on day 10 of the culture of the existing differentiation medium of porcine adipose-derived mesenchymal stem cells (positive control) and the modified differentiation medium of the present invention were measured by real-time fluorescence quantitative PCR (fig. 4, 5, and 6). The PPAR gamma gene is generally highly expressed in the early stage of differentiation, the differentiation capability of the adipose mesenchymal stem cells is represented, the expression level of FABP4 in the differentiation process is continuously increased, and the differentiation level of the adipose mesenchymal stem cells is represented.
The results showed that the relative expression level of PPAR γ gene was increased 1.49 (± 0.83) fold on day 5 of differentiation and 2.48 (± 0.75) fold on day 10 of differentiation, as compared to the old formulation. The relative expression of the Plin1 gene was increased 1.62 (. + -. 0.62) fold on day 10 of differentiation. Therefore, the improved cell differentiation medium of the present invention improves the differentiation capacity and differentiation level of adipose-derived mesenchymal stem cells.
2) Protein level:
according to the treatment method of example 1, on the 5 th and 10 th days of inducing differentiation in step 3), samples were taken, and the cells on the 5 th and 10 th days were cultured in the present differentiation medium for porcine adipose-derived mesenchymal stem cells (positive control) and the modified differentiation medium according to the present invention, respectively, in 2 plates, to which 100. Mu.L of RIPA (plus 1mM PMSF, final concentration) was added, and were lysed on ice for 30min and stored at-20 ℃ for further use. After centrifugation at 12000g for 5 minutes, the supernatant was collected, subjected to measurement and protein concentration using BCA kit from Sammerfer, and added with 5 Xloading buffer according to 4 (V: V), mixed well and heated at 95 ℃ for 5min to denature the protein, and stored at-80 ℃.
SDS-page gel electrophoresis, preparing electrophoresis buffer solution and transfer printing solution in advance (10% methanol is needed to be added to the transfer printing solution), under the condition that the electrophoresis buffer solution is not in a 12% denatured agarose precast gel plate, respectively adding denatured protein containing 20 mu g into an upper sample hole, setting voltage 80V electrophoresis for 30min, 120V electrophoresis for 90min and the two procedures above, and observing whether the protein loading solution reaches the bottom of the precast gel plate.
Film transfer: activating PVDF membrane in methanol for about 10s, storing in transfer solution, placing sponge, 2 layers of filter paper, gel, activated PVDF membrane, 2 layers of filter paper, and sponge, clamping with transfer clamp, placing in electrophoresis tank, adding prepared transfer solution, and running for 90min at 90V.
And (3) sealing: putting the PVDF membrane after the transfer printing into a sealing solution (5% skimmed milk powder prepared by TBST), sealing for 2h by a shaking table at room temperature, and then sucking off the sealing solution.
Primary antibody and secondary antibody incubation: PPAR gamma, FABP4 and Plin1 primary antibody were diluted as per the antibody specification and incubated at 4 ℃ for 14-16h. After the primary antibody incubation is finished, the primary antibody is recovered, and TBST is washed for three times, and each time is 5min. Diluted secondary antibody was added and incubated for 2h, and after completion TBST was washed three times for 5min each.
And (3) developing: and covering a PVDF membrane in a dark place by using a developing solution, incubating for 5min, then absorbing the developing solution, and taking a picture under a gel imager. And grey scale analysis was performed using Quantity One analysis software. The internal reference protein used in this experiment was Gapdh (fig. 7).
The results show that: compared with the traditional differentiation medium treatment group (positive control), the improved cell differentiation medium treatment group can improve the relative expression of the PPAR gamma protein by 1.49 (+ -0.83) times on the 5 th day of differentiation and improve the relative expression of the FABP4 protein by 1.97 (+ -0.06) times on the 10 th day of differentiation; the relative expression of the Plun 1 protein can be increased by 1.82 (+ -0.06) times at day 10 of differentiation. The improved cell differentiation culture medium has higher protein content of PPAR gamma, FABP4 and Plun 1 in induced differentiation.
The above disclosed examples are illustrative of the disclosed embodiments of the present invention and are not to be construed as limiting the invention, which is set forth in the following description of the preferred embodiments, and many variations and modifications may be made without departing from the scope and spirit of the invention. Indeed, various modifications of the above-described embodiments which are obvious to those skilled in the art to which the invention pertains are intended to be covered by the scope of the present invention.
Claims (9)
1. An improved cell differentiation medium for in vitro differentiation of a porcine adipose-derived mesenchymal stem cell is characterized in that the improved cell differentiation medium is a porcine adipose-derived mesenchymal stem cell differentiation medium added with an oleic acid oil component, and the improved cell differentiation medium can be safely used for food.
2. The improved cell differentiation medium according to claim 1, wherein said porcine adipose-derived mesenchymal stem cell differentiation medium comprises a porcine adipose-derived mesenchymal stem cell basal medium and a penicillin streptomycin double antibody solution, and the volume ratio of the porcine adipose-derived mesenchymal stem cell basal medium to the penicillin streptomycin double antibody solution is 99;
preferably, the basal medium of the porcine adipose-derived mesenchymal stem cells is selected from a DMEM medium, a MEM medium, a,
DMEM/F12 culture medium or F10 culture medium;
preferably, in the penicillin-streptomycin double-resistant solution, the content of penicillin is 10000U/ml, and the content of streptomycin is 10mg/ml.
3. The improved cell differentiation medium according to claim 1, wherein said cell culture accessory factors are selected from the group consisting of the following lipid differentiation promoting factors: insulin (Insulin), oil and fat, vitamin C, vitamin B, vitamin K and oleic acid.
4. The fat or oil according to claim 3, wherein the edible oil is commercially available, and the further edible oil is one selected from the group consisting of soybean oil, peanut oil, corn oil, sesame oil and olive oil, and is added at a concentration of 150 μ M to 350 μ M.
5. The modified cell differentiation medium according to claim 3, wherein any added lipotropic differentiation factor is present in a concentration of 0.5 μ g/mL to 0.2mg/mL.
6. The use of the improved cell differentiation medium of claim 1 for in vitro induction of differentiation of porcine adipose-derived mesenchymal stem cells, wherein the improved cell differentiation medium of claim 1 is used for in vitro induction of differentiation of porcine adipose-derived mesenchymal stem cells.
7. The use of claim 6, wherein the modified cell differentiation medium is capable of promoting differentiation of porcine adipose-derived mesenchymal stem cells during in vitro induced differentiation.
8. The use of claim 6, wherein the modified cell differentiation medium can increase the expression level of PPAR γ, FABP4 and Plin1 genes in the in vitro differentiation induction process of the porcine adipose-derived mesenchymal stem cells.
9. The use of claim 6, wherein the modified cell differentiation medium can increase the expression level of PPAR γ, FABP4 and Plun 1 proteins of pig adipose mesenchymal stem cells during in vitro induced differentiation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211286114.8A CN115896009A (en) | 2022-10-20 | 2022-10-20 | Efficient low-cost culture medium for in-vitro differentiation of adipose-derived mesenchymal stem cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211286114.8A CN115896009A (en) | 2022-10-20 | 2022-10-20 | Efficient low-cost culture medium for in-vitro differentiation of adipose-derived mesenchymal stem cells |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115896009A true CN115896009A (en) | 2023-04-04 |
Family
ID=86490494
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211286114.8A Pending CN115896009A (en) | 2022-10-20 | 2022-10-20 | Efficient low-cost culture medium for in-vitro differentiation of adipose-derived mesenchymal stem cells |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115896009A (en) |
-
2022
- 2022-10-20 CN CN202211286114.8A patent/CN115896009A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112708591B (en) | Culture medium with definite chemical components for in-vitro differentiation of muscle stem cells | |
| CN108330044B (en) | Method for resource utilization of yellow wine brewing water | |
| CN112592890A (en) | Method for promoting proliferation of muscle stem cells | |
| CN107535671B (en) | Microbial fermentation yellow wine lees feed for improving rumen protein utilization rate and preparation method thereof | |
| CN101892142A (en) | Preparation method of in-vivo pit skin mud | |
| CN112251379A (en) | Acid-resistant acetoin-producing Bacillus belgii DQA21 and application | |
| CN115404210B (en) | Method for inhibiting umbilical cord mesenchymal stem cell aging | |
| CN112553147A (en) | Growth factor composition for promoting proliferation of muscle stem cells and application thereof | |
| CN109266610A (en) | A method of promotion Derived from Mesenchymal Stem Cells is neuron | |
| CN115927172A (en) | Improved culture medium with specific chemical components for inducing in vitro adipogenic differentiation and application thereof | |
| CN115896009A (en) | Efficient low-cost culture medium for in-vitro differentiation of adipose-derived mesenchymal stem cells | |
| CN112899176B (en) | Grape saccharomyces cerevisiae capable of producing melatonin at high yield and application thereof | |
| CN111849880B (en) | Recovery method of human adipose mesenchymal stem cells after ultralow-temperature cryopreservation | |
| Kotarska et al. | Effect of various activators on the course of alcoholic fermentation | |
| CN104388384A (en) | Application of oxidized low density lipoprotein for inducing mesenchymal stem cell to differentiate to myocardium-like cell | |
| CN108034634B (en) | Method for separating endometrial mesenchymal stem cells from menstrual blood | |
| CN114752554B (en) | Serum-free myogenic differentiation medium containing natural compounds and application thereof | |
| CN106148277A (en) | A kind of mitochondria separation method being applicable to stem cell | |
| CN112877286B (en) | Stem cell in-vitro induction method | |
| CN110396489A (en) | A kind of artificial yeast for brewing rice wine leavening of yellow rice wine and its application method | |
| CN108828086A (en) | A kind of artificial bear gall powder and its method for evaluating quality | |
| CN111690598A (en) | Solen oligopeptide, and preparation method and application thereof | |
| CN110684721A (en) | Culture medium and method for evaluating embryo quality by using same | |
| CN110628546A (en) | Method for increasing content of tetramethylpyrazine in base liquor of Maotai-flavor liquor | |
| WO2024125632A1 (en) | Culture medium for inducing in-vitro adipogenic differentiation of cells and use thereof |
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 |