CN104711308A - Micromolecule substance for stimulating mesenchymal stem cells to secrete fibronectin - Google Patents
Micromolecule substance for stimulating mesenchymal stem cells to secrete fibronectin Download PDFInfo
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- CN104711308A CN104711308A CN201510163555.2A CN201510163555A CN104711308A CN 104711308 A CN104711308 A CN 104711308A CN 201510163555 A CN201510163555 A CN 201510163555A CN 104711308 A CN104711308 A CN 104711308A
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Abstract
The invention provides a micromolecule substance for stimulating mesenchymal stem cells to secrete fibronectin, which is named progestin and also called progesterone or pregnendione. The invention is characterized in that the molecular formula of the micromolecule substance is C21H30O2, and the molecular weight is 314.46. The molecular structure general formula of the micromolecule substance is disclosed in the specification. The micromolecule substance capable of stimulating cells to secrete adhesive molecules is used instead of the necessary and expensive fibronectin, thereby ensuring the adherent growth of the mesenchymal stem cells.
Description
Technical field
The invention belongs to the culture medium prescription composition in cell and tissue culture field, especially a kind of small-molecule substance stimulating mescenchymal stem cell Fibronectin Secretion.
Background technology
Mescenchymal stem cell, English name is mesenchymal stem cells (MSC), also other title is had, as marrow stromal cell, fat stem cell, pluripotency stroma cell etc., refer to that single or a group meets cell (the Dominici M of international uniform standard, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, ProckopDj, Horwitz E.Minimal criteria for defining multipotent mesenchymal stromalcells.The International Society for Cellular Therapy position statement.Cytotherapy, 2006, 8 (4): 315-7).Mescenchymal stem cell is present in Various Tissues and organ, comprise marrow, fat, umbilical cord, placenta, liver, brain, sclerotin etc., inside and outside all has the ability to the differentiation of bone, cartilage and fatty tissue, has the effects such as immunomodulatory, hematopoiesis support, angiogenesis promoting.
At present, multiple country has ratified autologous or allosome mescenchymal stem cell and has been applied to clinical, with the graft versus host disease (GVH disease) occurred in treatment and prevention hematopoietic stem cell transplantation process, cartilage injury, myocardial infarction and clone disease that treatment osteoarthritis merges merge the various diseases such as intestinal fistula.In addition, the clinical experimental study of mescenchymal stem cell deepens continuously, and the clinical trial of registration comprises treatment hepatic fibrosis, diabetes, peripheral vascular disease, necrosis of femoral head etc.But, for reaching treatment cell consumption, must amplification in vitro be carried out, and the system of amplification of mesenchymal stem cells mainly contains three kinds: (1) is containing the system of bovine serum; (2) containing the system of human blood goods; (3) serum-free system.The above two systems are cultivated mescenchymal stem cell and are used for the treatment of, not only patient will be exposed to suffer from the risk of zoonosis and transmissible disease, also because of transplanted cells death release foreign protein, organism immune response may be caused, causes serum sickness or other immunological disease.In addition, the activity of every serum batches and blood products is different, also have impact on the technical stability of amplifying cells.Therefore, in current clinical trial and application, the more employing serum free culture systems of people, carry out mescenchymal stem cell amplification.
Mescenchymal stem cell is a kind of adherent growth cell; For ensureing the effective amplification in vitro of cell, when using serum free culture system, the material promoting cell attachment must be added, as the material such as fibronectin (English name, fibronectin, abbreviation FN), vitronectin, collagen.Generally, the consumption of these materials is 5mg/L.Fibronectin is wherein most important molecule, is also the required composition of mesenchymal stem cell serum-free culture medium.But the firm quotes such as the material prices such as fibronectin are very expensive, Sigma are all more than 1mg/1000 Renminbi.Containing this composition in the human mesenchymal stem cell serum free medium that Stemcell company of Canada provides, this substratum total price is at 8000 yuans/more than 500ml.Adhesion molecule fibronectin should be the composition that substratum composition is more expensive.
Summary of the invention
For the weak point existed in the problems referred to above, the invention provides a kind of function utilizing this material incentive emiocytosis adhesion molecule, substitute the fibronectin of the cost intensive that must add, to ensure a kind of small-molecule substance stimulating mescenchymal stem cell Fibronectin Secretion of the adherent growth of mescenchymal stem cell.
For achieving the above object, the invention provides a kind of small-molecule substance stimulating mescenchymal stem cell Fibronectin Secretion, its name is called progesterone, also known as Progesterone or pregnendione, it is characterized in that, the molecular formula of this small-molecule substance is C21H30O2, and its molecular weight is 314.46;
The general formula of molecular structure of this small-molecule substance is:
Compared with prior art, the present invention has the following advantages:
The present invention utilizes small-molecule substance with the function of irritation cell secretion adhesion molecule, substitutes the fibronectin of the cost intensive that must add, to ensure the adherent growth of mescenchymal stem cell.
The small-molecule substance provided in the present invention can stimulate mescenchymal stem cell to synthesize fibronectin, thus increases the ability of cell adhesion at the bottom of culturing bottle/ware/plate.When using serum free medium to carry out derived mesenchymal stem cells in vitro amplification, in system, can not fibronectin be added, thus significantly reduce mesenchymal stem cell serum-free culture medium cost.In view of mescenchymal stem cell expresses PgR, Natural progesterone and synthetic progesterone can pass through cell surface receptor, cause cell that identical or approximate physiology change occurs.Therefore, indication progesterone of the present invention, also known as Progesterone, refers to a kind of natural progestogen secreted by corpus luteum, and also comprising with progesterone is the serial synthetic progestogen of molecular skeleton.
The present invention is particularly useful for when preparing mesenchymal stem cell serum-free culture system, reduces and adds fibronectin amount, or avoids adding fibronectin, thus reduces culture medium cost.But under low serum condition, can there is the physiological effect with serum-free condition lower aprons in cell surface receptor and ligand binding.Therefore, the present invention is still applicable to the preparation of the low blood serum medium of mescenchymal stem cell.In addition, current business-like mesenchyme serum free medium, is not only suitable for bone marrow mescenchymal stem cell and cultivates, and is also suitable for other derived mesenchymal stem cell and cultivates.Therefore, the present invention should be suitable for all tissue-derived mesenchymal stem cells, the cell cultivation process under low serum and serum-free condition.
The small-molecule substance provided in the present invention utilizes the biological function of culturing cell itself, is stimulated by allogenic material, for institute's culturing cell is adherent and propagation provides required nutritive ingredient.
Accompanying drawing explanation
Fig. 1 is under finite concentration Progesterone stimulated, human marrow mesenchymal stem cell form;
Fig. 2 is western blot results of hybridization, represents under finite concentration Progesterone stimulated, human marrow mesenchymal stem cell synthesis fibronectin situation;
Fig. 3 is cell adhesion experiments result, represents that, under finite concentration Progesterone stimulated, human marrow mesenchymal stem cell adheres to the ability of culture plate;
Fig. 4 is flow cyctometry analytical results, represents under the effect of finite concentration progesterone, human mesenchymal stem cell surface molecular expression;
Fig. 5 is MTT experiment result, represents under different concentrations of progesterone stimulates, Proliferation of Human Mesenchymal Stem Cells situation.
Embodiment
The invention provides a kind of small-molecule substance stimulating mescenchymal stem cell Fibronectin Secretion, its name is called progesterone, also known as Progesterone or pregnendione, it is characterized in that, the molecular formula of this small-molecule substance is C21H30O2, and its molecular weight is 314.46;
The general formula of molecular structure of this small-molecule substance is:
Experiment one, cellular form are observed
1, experiment material
(1) cell: human marrow mesenchymal stem cell, the third generation, derives from Healthy People, is our company's freeze-stored cell.
(2) progesterone: be Sigma Co., USA's product, article No. is P0130.Weighing pulvis is dissolved in DMSO, and 4 DEG C of packing store.
(3) ITS: be a kind of compound formulation, for serum replacement, is Sigma Products.Final concentration insulin-containing 10 μ g/ml, Transferrins,iron complexes 5.5 μ g/ml, Sodium Selenite 5ng/ml, linolic acid 4.70 μ g/ml, linolenic acid 4.70 μ g/ml, bovine serum albumin 500 μ g/ml.Article No. for the stoste that I2771.Sigma provides be 100X.
(4) alpha improves bottom line basic medium (alpha minimum essential medium, alpha-MEM), is a kind of basic medium cultivated mescenchymal stem cell and commonly use.For Sigma Products, article No. is M0894.
(5) l-GLUTAMINE (L-glutamine) is sigma product, article No. G6392.Use aseptic double-distilled water dissolves, and total concn is 2mM.
2, experimental technique
After human marrow mesenchymal stem cell recovery, alpha-MEM washing once.According to 5 × 10
4/ ml is suspended in the alpha-MEM substratum containing ITS and l-GLUTAMINE, is inoculated in 100mm culture dish.Add progesterone 25ng/ml in system, not add progesterone cell for contrast, cultivate and observe under inverted microscope after 72 hours, take a picture.
3, experimental result
As shown in Figure 1, two groups of cellular fories, all in inoblast sample, meet typical mescenchymal stem cell morphological feature (Fig. 1) to result.PRELIMINARY RESULTS is seen, in the progesterone treatment group visual field, cell is more.
After human mesenchymal stem cell inoculates 72 hours, do not add progesterone group (left side) and add progesterone group (right side) cell all in inoblast sample.Indicate: 50 μm.
Experiment two, western blot hybridization
1, test materials
(1) cell: human marrow mesenchymal stem cell 2 strain, the third generation, derives from two Healthy Peoples respectively, is our company's freeze-stored cell.
(2) progesterone: be Sigma Co., USA's product, article No. is P0130.Weighing pulvis is dissolved in DMSO, and 4 DEG C of packing store.
(3) ITS: be a kind of compound formulation, for serum replacement, is Sigma Products.Final concentration insulin-containing 10 μ g/ml, Transferrins,iron complexes 5.5 μ g/ml, Sodium Selenite 5ng/ml, linolic acid 4.70 μ g/ml, linolenic acid 4.70 μ g/ml, bovine serum albumin 500 μ g/ml.Article No. for the stoste that I2771.Sigma provides be 100X.
(4) alpha improves bottom line basic medium (alpha minimum essential medium, alpha-MEM), is a kind of basic medium cultivated mescenchymal stem cell and commonly use.For Sigma Products, article No. is M0894.
(5) antibody: the goat anti-mouse antibody of mouse anti human FN antibody, mouse anti human GAPDH antibody, horseradish peroxidase-labeled, all purchased from Santa Cruz company.
(6) enhanced chemical luminescence reagent is ECL product.
2, test method
Utilize the system cultivator marrow MSC of the alpha-MEM containing ITS and l-GLUTAMINE, 72 h before harvest cells, are suspended in above-mentioned culture system, are inoculated in 100mm culture dish.Add 25ng/ml progesterone, continue to cultivate cultivation after 72 hours, tryptic digestion harvested cell.Utilize the RIPA lysis buffer lysing cell containing 1mM PMSF, 5mM EDTA and 1mM inhibitors of phosphatases, collected by centrifugation supernatant, BCA method measures protein concentration.According to 40 μ g/ road loadings, albumen, after 6% polyacrylamide gel electrophoresis is separated, is transferred on nitrocellulose filter.Utilize the TBS buffer blind containing 1%BSA to spend the night, add anti-FN and anti-GAPDH antibody, antibody concentration is 1: 1000.Room temperature reaction is after 2 hours, and TBS buffer solution 5 times, adds the anti-mouse antibody of horseradish peroxidase-labeled, room temperature reaction 30 minutes, chemoluminescence method display result.
3, test-results
As shown in Figure 2, after Progesterone stimulated, in cell, fibronectin content is apparently higher than control group, and it is close respectively to organize GAPDH ribbon density for result.
Human marrow mesenchymal stem cell is through Progesterone stimulated after 72 hours, and lysing cell take GAPDH as internal reference, western blot hybridization check cell FN content.1-2 road: do not add progesterone cell for contrast (control); 3-4 road: progesterone process cell (progesterone).
Experiment three, cell adhesion experiments
1. material and reagent
(1) cell: human marrow mesenchymal stem cell 3 strain, the third generation, derives from three Healthy Peoples respectively, is our company's freeze-stored cell.
(2) progesterone: be Sigma Co., USA's product, article No. is P0130.Weighing pulvis is dissolved in DMSO, and 4 DEG C of packing store.
(3) ITS: be a kind of compound formulation, for serum replacement, is Sigma Products.Final concentration insulin-containing 10 μ g/ml, Transferrins,iron complexes 5.5 μ g/ml, Sodium Selenite 5ng/ml, linolic acid 4.70 μ g/ml, linolenic acid 4.70 μ g/ml, bovine serum albumin 500 μ g/ml.Article No. for the stoste that I2771.Sigma provides be 100X.
(4) alpha improves bottom line basic medium (alpha minimum essential medium, alpha-MEM), is a kind of basic medium cultivated mescenchymal stem cell and commonly use.For Sigma Products, article No. is M0894.
(5) MTT and DMSO is Sigma Products.
2, experimental technique
Collect the compared with control cells stimulating people's marrow MSC of 72 hours through progesterone (25ng/ml) and do not add progesterone cultivation, be resuspended in respectively in the α-MEM substratum of ITS and l-GLUTAMINE, and to adjust cell concn be 1 × 10
6/ ml.Add cell suspension to 96 orifice plates by every hole 36 μ l, supply volume to 100 μ l.Cultivate after 2 hours for 37 DEG C, remove non-attached cell and substratum, PBS washs 2 times.Add MTT reagent, final concentration reaches 100 μ g/ml, and 37 DEG C are continued to hatch 4 hours.Add DMSO100 μ l/ hole, 490nm wavelength measures absorbancy, according to OD value display attached cell quantity.
3. experimental result
Utilize the MSC experiment prompting of 3 sample sources, untreated cell OD490 mean value is respectively 0.23 ± 0.04,0.19 ± 0.01,0.22 ± 0.01, significantly lower than progesterone treatment group cell (p value is all less than 0.01), the latter OD490 mean value is respectively 0.29 ± 0.01,0.23 ± 0.01,0.27 ± 0.03.
Result as shown in Figure 3.Three strain human marrow mesenchymal stem cells are after progesterone (25ng/ml) stimulates 72 hours, and collecting cell, plants in 96 orifice plates, cultivate 2 hours.MTT experiment observes adherent cell quantity.Ordinate zou: OD490, represents cell relative populations; X-coordinate: from the result of three samples.
Experiment four, cells characteristic surface molecular detect
1. material and reagent
(1) cell: human marrow mesenchymal stem cell 2 strain, the third generation, derives from two Healthy Peoples respectively, is our company's freeze-stored cell.
(2) progesterone: be Sigma Co., USA's product, article No. is P0130.Weighing pulvis is dissolved in DMSO, and 4 DEG C of packing store.
(3) ITS: be a kind of compound formulation, for serum replacement, is Sigma Products.Final concentration insulin-containing 10 μ g/ml, Transferrins,iron complexes 5.5 μ g/ml, Sodium Selenite 5ng/ml, linolic acid 4.70 μ g/ml, linolenic acid 4.70 μ g/ml, bovine serum albumin 500 μ g/ml.Article No. for the stoste that I2771.Sigma provides be 100X.
(4) alpha improves bottom line basic medium (alpha minimum essential medium, alpha-MEM), is a kind of basic medium cultivated mescenchymal stem cell and commonly use.For Sigma Products, article No. is M0894.
(5) the mouse anti human CD31 that antibody: PE marks, CD45, CD73 and CD90 monoclonal antibody is BD Products.CD31 is endothelial cell characteristics mark; CD45 is hematopoietic cell markers; CD73 is mesenchymal cell mark; CD90 is often expressed in stem cell surface.
2, experimental technique
Human marrow mesenchymal stem cell is suspended in the α-MEM substratum containing ITS and l-GLUTAMINE, adds progesterone (25ng/ml) and cultivate 72 h before harvest cells.After PBS washes twice, add above-mentioned antibody respectively, room temperature lucifuge reacts 20 minutes.After PBS washes twice, FACSCalibur collecting cell, 10000 point data are at least collected in each reaction.WinMdi2.9 software analysis results data, establish a removal cell debris in analytic process.
3. experimental result
Result as shown in Figure 4, human marrow mesenchymal stem cell is cultivated in above-mentioned system, a cell surface minute feature meets mescenchymal stem cell feature, namely do not express CD31 (endothelial cell marker) and CD45 (hemocyte mark), express CD73 (mesenchymal cell mark) and CD90 (stem cell labeling).Result shows the cell after Progesterone stimulated, the change of phenotype do not detected.
Derive from two Healthy People mesenchymal stem cells MSCs, after progesterone (25ng/ml) stimulates 72 hours, collecting cell, carries out antibody labeling.Ordinate zou: number of data points; X-coordinate: relative intensity of fluorescence.
Test five, MTT experiment detect ability of cell proliferation
1. material and reagent
(1) cell: human marrow mesenchymal stem cell 1 strain, the third generation, derives from Healthy People, is our company's freeze-stored cell.
(2) progesterone: be Sigma Co., USA's product, article No. is P0130.Weighing pulvis is dissolved in DMSO, and 4 DEG C of packing store.
(3) ITS: be a kind of compound formulation, for serum replacement, is Sigma Products.Final concentration insulin-containing 10 μ g/ml, Transferrins,iron complexes 5.5 μ g/ml, Sodium Selenite 5ng/ml, linolic acid 4.70 μ g/ml, linolenic acid 4.70 μ g/ml, bovine serum albumin 500 μ g/ml.Article No. for the stoste that I2771.Sigma provides be 100X.
(4) alpha improves bottom line basic medium (alpha minimum essential medium, alpha-MEM), is a kind of basic medium cultivated mescenchymal stem cell and commonly use.For Sigma Products, article No. is M0894.
(5) MTT and DMSO is Sigma Products.
(6) Prostatropin (english abbreviation: bFGF) is purchased from PeproTech company, and final concentration is 10ng/ml.
2. experimental technique
Collector's marrow MSC, by 1.0 × 10
6/ ml is suspended in the alpha-MEM containing ITS and glutamine, is inoculated in by cell in 100mm culture dish, cultivates trypsinization collecting cell after 72 hours.Centrifuge washing, meter cell count, and cell suspension is transferred to 96 orifice plates, every hole is containing 3600 cells, cumulative volume is 100 μ l, is respectively 0,5ng/ml in system containing final concentration, 10ng/ml, the progesterone of 25ng/ml, 50ng/ml, 100ng/ml, or the Prostatropin of 10ng/ml (basicfibroblast growth factor, bFGF).Cell is placed in 37 DEG C, cultivate 72 hours under 5%CO2 condition, every hole adds MTT (1mg/ml) 10 μ l, and continue cultivation after 4 hours, remove culture supernatant, every hole adds dimethyl sulfoxide (DMSO) 100 μ l.Enzyme linked immunological instrument measures every hole optical density value, mensuration wavelength is 490nm.
3, experimental result
Utilize serum-free system cultivator marrow MSC, add the progesterone of different concns, not add progesterone group for negative control, to add bFGF group for positive control, after 72 hours, MTT experiment measures each group of cell viability.Result is as shown in Figure 5: adding bFGF (10ng/ml) in (1) system obviously can promote cell proliferation, OD490 mean value is 0.42 ± 0.04, be significantly higher than negative control group (0.29 ± 0.02, p < 0.001), consistent with expected results, bFGF group can be used as positive control; (2) progesterone (5-50ng/ml) within the scope of finite concentration, MSC propagation is had no effect, but when concentration is increased to 100ng/ml, cell proliferation rate slows down, OD490 mean value is 0.26 ± 0.02, significantly lower than negative control group (p < 0.05).Result is pointed out, and using progesterone composition in MSC culture system, it is suitable for selecting final concentration to be 25ng/ml.
Ctr: the mescenchymal stem cell not adding progesterone; BFGF: add Prostatropin as positive control.
The foregoing is only preferred embodiment of the present invention, is only illustrative for invention, and nonrestrictive.Those skilled in the art is understood, and can carry out many changes in the spirit and scope that invention claim limits to it, amendment, even equivalence, but all will fall within the scope of protection of the present invention.
Claims (1)
1. stimulate a small-molecule substance for mescenchymal stem cell Fibronectin Secretion, its name is called progesterone, also known as Progesterone or pregnendione, it is characterized in that, the molecular formula of this small-molecule substance is C21H30O2, and its molecular weight is 314.46;
The general formula of molecular structure of this small-molecule substance is:
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105002241A (en) * | 2015-07-08 | 2015-10-28 | 河南中科干细胞基因工程有限公司 | Application of progesterone in stimulation of fibronectin secretion of mesenchymal stem cells |
| CN106479971A (en) * | 2016-12-28 | 2017-03-08 | 深圳江淼医疗有限公司 | A kind of serum-free medium for cultivating mescenchymal stem cell and method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102634500A (en) * | 2011-05-25 | 2012-08-15 | 侯亚义 | Genetic engineering application of quick decidua and placenta mesenchymal stem cells (MSC) obtaining and micro-molecular modifying |
| CN102827807A (en) * | 2012-09-19 | 2012-12-19 | 北京京蒙高科干细胞技术有限公司 | Serum-free culture medium for mesenchymal stem cells |
| CN102936578A (en) * | 2012-11-12 | 2013-02-20 | 山东省齐鲁干细胞工程有限公司 | Method for strengthening immune-suppression function of mesenchymal stem cells |
| CN103881972A (en) * | 2013-08-28 | 2014-06-25 | 中国人民解放军南京军区福州总医院 | Method applicable to serum-free culture of mesenchymal stem cells |
-
2015
- 2015-04-09 CN CN201510163555.2A patent/CN104711308A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102634500A (en) * | 2011-05-25 | 2012-08-15 | 侯亚义 | Genetic engineering application of quick decidua and placenta mesenchymal stem cells (MSC) obtaining and micro-molecular modifying |
| CN102827807A (en) * | 2012-09-19 | 2012-12-19 | 北京京蒙高科干细胞技术有限公司 | Serum-free culture medium for mesenchymal stem cells |
| CN102936578A (en) * | 2012-11-12 | 2013-02-20 | 山东省齐鲁干细胞工程有限公司 | Method for strengthening immune-suppression function of mesenchymal stem cells |
| CN103881972A (en) * | 2013-08-28 | 2014-06-25 | 中国人民解放军南京军区福州总医院 | Method applicable to serum-free culture of mesenchymal stem cells |
Non-Patent Citations (4)
| Title |
|---|
| HUI HUI ZHU等: "Progestin stimulates the biosynthesis of fibronectin and accumulation of fibronectn mRNA in human endometrial stromal cells", 《HUMAN REPRODUCTION》 * |
| JIN CHEN等: "Thrombin promotes fibronectin secretion by bone marrow mesenchymal stem cells via the protease-activated receptor mediated signalling pathways", 《STEM CELL RESEARCH & THERAPY》 * |
| 周宁波等主编: "《化学工业出版社》", 30 August 2010 * |
| 王利公等: "子宫内膜基质干细胞的分离培养研究", 《浙江大学学报(医学版)》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105002241A (en) * | 2015-07-08 | 2015-10-28 | 河南中科干细胞基因工程有限公司 | Application of progesterone in stimulation of fibronectin secretion of mesenchymal stem cells |
| CN106479971A (en) * | 2016-12-28 | 2017-03-08 | 深圳江淼医疗有限公司 | A kind of serum-free medium for cultivating mescenchymal stem cell and method |
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Application publication date: 20150617 |