CN105087479A - Stem cell serum-free culture medium and stem cell culture method - Google Patents
Stem cell serum-free culture medium and stem cell culture method Download PDFInfo
- Publication number
- CN105087479A CN105087479A CN201510518943.8A CN201510518943A CN105087479A CN 105087479 A CN105087479 A CN 105087479A CN 201510518943 A CN201510518943 A CN 201510518943A CN 105087479 A CN105087479 A CN 105087479A
- Authority
- CN
- China
- Prior art keywords
- stem cell
- culture medium
- serum
- free culture
- concentration
- 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
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a stem cell serum-free culture medium. The stem cell serum-free culture medium comprises a basal culture medium, insulin, glutamine, ferruginous food additives, sodium selenite, progestone and putrescine. The invention also discloses a stem cell culture method. The stem cell culture method comprises the following steps of acquiring stem cells; and culturing the stem cells in vitro by using the stem cell serum-free culture medium. Through cells obtained by the stem cell serum-free culture medium, the problem of cell purification difficulty caused by pollution of pathogens carried by a serum culture medium can be avoided, stability of cell production can be improved, cell proliferation quantity is increased, and cell viability is improved.
Description
Technical field
The present invention relates to tissue engineering technique field, particularly the cultural method of stem cell serum-free culture medium and stem cell.
Background technology
At present, in the cellar culture system of stem cell, all add a certain proportion of animal serum, relatively more conventional is foetal calf serum or newborn calf serum.The very complicated mixture that serum is made up of the biomolecules that much varies in size, Main Function when it is cultivated in vitro to cell is to provide somatomedin, hormone, associated proteins, and provides provide protection.But it is also unfavorable for supressor or the toxicant of Growth of Cells containing some, have potential cytotoxic effect.In serum, the protein of a large amount of complicated component, brings difficulty to the stdn of cell cultures, also expresses separation and purification of products to cell cultures simultaneously and brings very large difficulty.And may there is the known or unknown pathogenic agent that animal carries in animal serum, constitute a threat to, add difficulty to for pilot scale culture cell to later possible clinical application.
Summary of the invention
Technical problem to be solved by this invention is, for utilizing animal serum culturing stem cells to there is the defects such as certain toxic action and pathogen contamination in prior art, provides a kind of nontoxicity and free of contamination stem cell serum-free culture medium.
The technical solution adopted for the present invention to solve the technical problems is: a kind of stem cell serum-free culture medium, comprises basic medium, also comprises Regular Insulin, glutamine, the foodstuff additive of iron content and Sodium Selenite, progesterone and butanediamine.
In stem cell serum-free culture medium provided by the invention, the foodstuff additive of described iron content to comprise in Transferrins,iron complexes, ironic citrate, ferric cacodylate and Gluconate Ferrecex one or more.
In stem cell serum-free culture medium provided by the invention, the concentration that in described stem cell serum-free culture medium, the concentration of Regular Insulin is 1-15 μ g/ml, the concentration of glutamine is 1-5mmol/l, the concentration of the foodstuff additive of iron content is 20-200ng/ml, the concentration of Sodium Selenite is 20-50nmol/l, the concentration of progesterone is 5-30mmol/l and butanediamine is 10-100 μm of ol/l.
In stem cell serum-free culture medium provided by the invention, also comprise and also comprise HEPES and penicillin.
In stem cell serum-free culture medium provided by the invention, the concentration of described HEPES is 2-20mmol/l, and the concentration of penicillin is 10-100mg/ml.
In stem cell serum-free culture medium provided by the invention, also comprise glucose, NaHCO
3, soybean pancreatin inhibitor, linolic acid and Streptomycin sulphate.
In stem cell serum-free culture medium provided by the invention, the massfraction of described glucose is 0.1-1%, NaHCO
3concentration be 1-10mmol/l, the massfraction of soybean pancreatin inhibitor is 0.05-0.2%, and linoleic concentration is 10-100 μm of ol/l, and the concentration of Streptomycin sulphate is 10-100mg/ml.
In stem cell serum-free culture medium provided by the invention, described basic medium is DMEM/F12 substratum.
The present invention further provides the cultural method of a kind of stem cell, comprise the steps:
Obtain stem cell, adopt above-mentioned stem cell serum-free culture medium to carry out vitro culture.
Implement the cultural method of stem cell serum-free culture medium provided by the invention and stem cell, following beneficial effect can be reached: 1, employing stem cell serum-free culture medium can avoid the mass discrepancy in prior art between serum batch, improves the repeatability of cell cultures and test-results; The cytotoxic effect of prior art 2, can be avoided to exist exogenous pollution that serum brings and serum component; 3, the serum component failed to understand can be avoided the impact of cell cultures and experimental study; 4, relatively clear and definite, the uniform quality of composition and protein content is low, is conducive to improving stability that cellular product produces and makes cellular product be easy to purifying; 5, the GI phase of cell can be extended or force cell to be in the maintenance high cell densities cultivation of G0 phase and long period, thus the product of production object as much as possible, and keep the characteristic of stem cell better.
Embodiment
The existence of the culture medium culturing stem cell serum component containing animal serum is adopted easily to produce toxic action for solving in prior art, and carry pathogenic agent and the defects such as cellular segregation purification difficult, innovative point of the present invention is to provide a kind of stem cell serum-free culture medium, can the recruitment factor of alternative serum by adding in the medium, thus achieve pollution-free culturing stem cells, improve the object of cellular product production stability.
Stem cell serum-free culture medium provided by the invention, comprises basic medium, Regular Insulin, glutamine, the foodstuff additive of iron content, Sodium Selenite, progesterone and butanediamine.
Wherein, glucose in basic medium is the significant energy source in growth and proliferation of cell and Product Expression process, preferably, basic medium is DMEM/F12 substratum, containing the extremely abundant and complicated nutritive substance needed for Growth of Cells in this DMEM/F12 substratum, can support that broad variety cell grows in the substratum of serum-free.
Regular Insulin can promote cells use glucose and protein, and promote the synthesis of RNA, protein and lipid acid, inhibited apoptosis is very important liability factor; Preferably, in stem cell serum-free culture medium, the concentration of Regular Insulin is 1-15 μ g/ml.
After glutamine desamidizate, can be used as the energy derive of culturing cell, and participate in synthesis and the nucleic acid metabolism of protein, promote the growing multiplication of cell; Preferably, the concentration of glutamine is 1-5mmol/l.
Trace elements iron is that growth and proliferation of cell is necessary, and the foodstuff additive of iron content can be one or more in Transferrins,iron complexes, ironic citrate, ferric cacodylate and Gluconate Ferrecex; Preferably, the foodstuff additive of iron content are Transferrins,iron complexes, because all there is specific TfR in most of mammalian cell, it is the main source that cell obtains required trace elements iron that TfR is combined with transferrin complex of protein, in addition, Transferrins,iron complexes also have somatomedin character and can with other trace element as combinations such as vanadium, for growth and proliferation of cell provides required trace element.Preferably, in stem cell serum-free culture medium, the concentration of the foodstuff additive of iron content is 20-200ng/ml.
Further, in cell cultivation process, tyrosine in basic medium, tryptophane, riboflavin, ascorbate salt etc. can produce a large amount of hydrogen peroxide, hydrogen peroxide is the major toxicity material of Growth of Cells and clone, and the saturated fatty acid on cytolemma, cell protein can be caused to be oxidized and/or DNA damage and cause necrocytosis.Therefore, appropriate Sodium Selenite is added in stem cell serum-free culture medium provided by the invention, trace element-selenium in Sodium Selenite participates in the mechanism of Selenoperoxidase and superoxide dismutase, thus can eliminate oxide compound enzyme and oxyradical to the injury of cell; Preferably, in this stem cell serum-free culture medium, the concentration of Sodium Selenite is 5-20nmol/l.
Progesterone can the propagation of irritation cell, and can promote cell migration, avoid cell attachment to grow problem that the cellular portions caused cannot touch substratum; Preferably, the concentration of progesterone is 5-30mmol/l.
Butanediamine, as the trace element in cell cultures and lower molecular weight nutritional factor, is the lipid needed for cytolemma synthesis and the water-soluble lipid needed for Growth of Cells, can promotes the growing multiplication of cell; Preferably, the concentration of butanediamine is 10-100 μm of ol/l.
In sum, the pathogen contamination that stem cell serum-free culture medium provided by the invention not only can avoid blood serum medium culturing cell to cause, and the problem such as cellular segregation purifying, and the growing multiplication of cell can be promoted.
Further, stem cell serum-free culture medium provided by the invention also comprises penicillin, and HEPES (Chinese name: hydroxyethyl piperazine second thiosulfonic acid, 2-[4-(2-Hydroxyethyl)-1-piperazinyl] ethanesulfonicacid,), these materials all have for the growing multiplication of cell and promote booster action energetically.
Wherein, HEPES can compensate the surge capability that in substratum, serum-free causes and decline, and to maintain the potential of hydrogen needed for Growth of Cells, preferably, the concentration of HEPES is 2-20mmol/l; Penicillin can hinder the Cell wall synthesis of bacterium, causes cellular leakage dead, thus the growth of anti-bacteria, preferably, the concentration of penicillin is 10-100mg/ml.
Further, stem cell serum-free culture medium provided by the invention also comprises glucose, NaHCO
3, soybean pancreatin inhibitor, linolic acid and Streptomycin sulphate.Wherein, glucose is the significant energy source in growth and proliferation of cell and Product Expression process, and preferably, the massfraction of glucose is 0.1-1%.NaHCO
3for be placed in when cell incubator cultivate time, regulate the pH value of substratum to 7.2-7.4, preferably, NaHCO
3concentration be 1-10mmol/l.Soybean pancreatin inhibitor can stop trypsin digestion and cell in passage process, reaches the object of Cell protection, and preferably, the massfraction of soybean pancreatin inhibitor is 0.05-0.2%.Linolic acid, as the trace element in cell cultures and lower molecular weight nutritional factor, is the lipid needed for cytolemma synthesis and the water-soluble lipid needed for Growth of Cells, Promote cell's growth; Preferably, linoleic concentration is 10-100 μm of ol/l.Streptomycin sulphate can act on the rrna of bacterium, and impede protein is translated, thus bacteria growing inhibiting; Under physical environment, do not consider the artificial resistance produced, responsive to Streptomycin sulphate to the insensitive most microorganism of penicillin, vice versa; Therefore, in the present invention, penicillin and Streptomycin sulphate collocation use can be controlled almost all common bacterium; Preferably, the concentration of Streptomycin sulphate is 10-100mg/ml.
In addition, many cells must adherently could grow, and in stem cell serum-free culture medium, owing to lacking the various adhesion anchoring factors in serum, cell often with suspension form growth, and is unfavorable for cell proliferation.Therefore, this defect is made up by adding short adherent material in stem cell serum-free culture medium, short adherent material is generally extracellular matrix, can be fibronectin or ln etc., simultaneously, the differentiation factor of short adherent material or important mitogen and maintenance normal cell function, to cellulous breeding and differentiation play an important role perhaps.Preferably, short adherent material is fibronectin, main promote mesoblastemic adherent with differentiation; In stem cell serum-free culture medium, fibronectin or or the concentration of ln be 0.1-2 μ g/ml, specifically can according to the adherent property of different cell, selectivity is added.
The present invention further provides the cultural method of a kind of stem cell, in the present embodiment, for mescenchymal stem cell, certain serum free medium of the present invention also can be applicable to the stem cell of other types, and the cultural method of the present embodiment mescenchymal stem cell comprises the steps:
1) mescenchymal stem cell is obtained;
2) stem cell serum-free culture medium provided by the invention is adopted to carry out amplification in vitro cultivation.
Wherein, step 1) obtain the process of mescenchymal stem cell and be:
Get people's bones of the body joint or Lower limb bone bone marrow prepare, anticoagulant heparin, be added in the centrifugal 30min of 2300rpm on peroll parting liquid that density is 1.073g/ml in the ratio of 1:2, get middle level monocyte, add PBS damping fluid, the centrifugal 10min of 1000rpm, wash 3 times, abandon supernatant liquor.The primary mescenchymal stem cell of gained counts under an optical microscope, is adjusted to 1 × 10
7individual/culturing bottle (25ml) density DMEM/F12 substratum is cultivated.
Be understandable that, these are only wherein a kind of approach obtaining mescenchymal stem cell provided by the invention, in prior art, the obtain manner of mescenchymal stem cell is equally applicable to the present invention.
Step 2) detailed process is:
When step 1) in Growth of Cells to when almost merging completely, incline old substratum, with PBS liquid rinsing 2 ~ 3 times, incline PBS liquid, the trypsin adding 0.25% includes 0.02%EDTA ethylenediamine tetraacetic acid (EDTA)) digestion, add-on is as the criterion with the cell at the bottom of making trypsinase can cover completely bottle.Observe under culturing bottle being placed in inverted microscope, see that kytoplasm bounces back, after intercellular substance increases, add stem cell serum-free culture medium provided by the invention, stop digestion, suction pipe order is blown and beaten at the bottom of bottle gently, under inverted microscope after the completely de-wall of observation of cell, go down to posterity in 1:2 ~ 1:3 ratio and be inoculated in 25ml culturing bottle, be positioned over 37 DEG C, 5%CO
2, 95% humidity incubator in cultivate, until Growth of Cells is to when almost merging completely, again go down to posterity, every 2-3 days changes liquid once.
In order to make object of the present invention, technical scheme and advantage clearly understand, cultivating the embodiment of mescenchymal stem cell below in conjunction with different dry cell non-serum culture medium provided by the invention, 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
In this embodiment, the component of stem cell serum-free culture medium and the concentration of each component in stem cell serum-free culture medium are respectively:
DMEM/F12 substratum: 100mL;
Sodium Selenite: 50 μ g/ml;
Transferrins,iron complexes: 150 μ g/ml;
Ironic citrate: 50 μ g/ml;
Regular Insulin: 15 μ g/ml;
The concentration of glutamine is 5mmol/l;
The concentration of progesterone is 10mmol/l;
The concentration of butanediamine is 40 μm of ol/l.
Embodiment 2
In this embodiment, the component of stem cell serum-free culture medium and the concentration of each component in stem cell serum-free culture medium are respectively:
DMEM/F12 substratum: 500mL;
Sodium Selenite: 40 μ g/ml;
Transferrins,iron complexes: 100 μ g/ml;
Regular Insulin: 7 μ g/ml;
The concentration of glutamine is 3mmol/l;
The concentration of HEPES is 10mmol/l;
The concentration of progesterone is 15mmol/l;
The concentration of butanediamine is 60 μm of ol/l;
The concentration of penicillin is 60mg/ml.
Embodiment 3
In this embodiment, the component of stem cell serum-free culture medium and the concentration of each component in stem cell serum-free culture medium are respectively:
Sodium Selenite: 30 μ g/ml;
Transferrins,iron complexes: 100 μ g/ml;
Regular Insulin: 2.5 μ g/ml;
The concentration of glutamine is 2mmol/l;
The concentration of HEPES is 5mmol/l;
The concentration of progesterone is 20mmol/l;
The concentration of butanediamine is 60 μm of ol/l;
The concentration of penicillin is 50mg/ml;
The massfraction of glucose is 0.6%;
NaHCO
3concentration is 3mmol/l;
The massfraction of soybean pancreatin inhibitor is 0.1%;
Linolic acid 60 μm of ol/l;
The concentration of Streptomycin sulphate is 50mg/ml.
For verifying that stem cell serum-free culture medium provided by the invention has outstanding significance effect further, further describe below by way of specific experiment.
Detected object:
Experimental group 1-3: the corresponding stem cell serum-free culture medium utilizing embodiment of the present invention 1-3 to provide is according to above-mentioned steps 2 respectively) method steps cultivate the cell obtained;
Control group: adopt and replace the stem cell serum-free culture medium in the present invention to cultivate the cell obtained containing the foetal calf serum of 10% (WT), the DMEM high glucose medium of l0mmol/lHEPES, l00U/ml mycillin; Wherein, the step adopting NaOH solution adjustment substratum pH value to 7.2-7.4 is also comprised.
One, test experience one
Mtt assay detects cytoactive
Get 5 piece of 96 orifice plate, every block 96 orifice plate is with 5 × 10
3the density in/hole experimental group and control group are cultivated obtain the 3rd generation cell be respectively inoculated in 5 orifice plates, the every hole of control group adds the low sugar DMEM liquid 200 μ l containing 10% (WT) foetal calf serum, the every hole of experimental group 1-3 adds the stem cell serum-free culture medium 200 μ l corresponding with embodiment 1-3 respectively, only add respectively in the hole not having cell in addition containing the low sugar DMEM substratum of 10% (WT) foetal calf serum and self-control stem cell serum-free culture medium each 200 μ L as blank, each 5 holes, be placed in 37 DEG C, 5%CO
2cultivate in incubator, experimental port and control wells change liquid once in every three days.
After this every day same time, randomly draw a plate, measure photoabsorption (OD) value.During measurement, every hole adds the 5mg/mlMTT solution of 20 μ L, continues at 37 DEG C, 5%CO
2cultivate in incubator after 6 hours, careful suction abandons liquid in hole, adds the DMSO (dimethyl sulfoxide (DMSO)) of 150 μ L in every hole.96 orifice plates are placed on enzyme-linked immunosorbent assay instrument and shake 20 minutes, adopt dual wavelength method, enzyme-linked immunoassay instrument setting determined wavelength 492nm, reference wavelength 630nm, measure OD value, the OD value according to recording judges viable cell quantity, OD value is larger, and cytoactive is stronger.
Table 1:
| Number of days | Experimental group 1 | Experimental group 2 | Experimental group 3 | Control group | Blank group |
| 1 | 0.0565±0.0218 | 0.0567±0.0228 | 0.0568±0.0225 | 0.0588±0.0151 | 0.0142±0.0003 |
| 2 | 0.0810±0.0181 | 0.0822±0.0187 | 0.0839±0.0189 | 0.0779±0.0129 | 0.0168±0.0023 |
| 3 | 0.1162±0.0212 | 0.1227±0.0222 | 0.1269±0.0226 | 0.0785±0.0230 | 0.0152±0.0057 |
| 4 | 0.1512±0.0211 | 0.1677±0.0214 | 0.1715±0.0215 | 0.1379±0.0188 | 0.0157±0.0021 |
| 5 | 0.2573±0.0160 | 0.3019±0.0167 | 0.3077±0.0169 | 0.2017±0.0177 | 0.0138±0.0033 |
| 6 | 0.3571±0.0131 | 0.4566±0.0133 | 0.4575±0.0136 | 0.2929±0.0234 | 0.0141±0.0085 |
| 7 | 0.6091±0.0148 | 0.8077±0.0151 | 0.8093±0.0154 | 0.4466±0.0281 | 0.0147±0.0027 |
| 8 | 1.6592±0.0159 | 1.6053±0.0161 | 1.6358±0.0162 | 0.7350±0.0131 | 0.0120±0.0051 |
| 9 | 1.8126±0.0169 | 1.8173±0.0170 | 1.8219±0.0171 | 0.7837±0.0181 | 0.0533±0.0044 |
| 14 | 1.7928±0.0118 | 1.8015±0.0122 | 1.8121±0.0123 | 0.9229±0.0128 | 0.0567±0.0033 |
Detected result: from the data in table 1, the OD value of experimental group 1-3 is all greater than control group, it can thus be appreciated that the mescenchymal stem cell quantity obtained by stem cell serum-free culture medium provided by the invention is more, and viable cell quantity is more, and activity is higher.
Test experience two, cell cycle are detected
Get experimental group respectively and control group cultivates the cell obtained for the 14th day, through flow cytometry G0/G1 phase, G2/M phase cell percentages, respectively get 5 groups of samples respectively and carry out detection and analyze.
Table 2:
Detected result: as shown in Table 2, in experimental group 1-3, G
0/ G
1phase and G
2/ M phase cell percentages is all greater than control group; It can thus be appreciated that, the viable cell quantity that the viable cell quantity that experimental group 1-3 is obtained by stem cell serum-free culture medium is obtained by blood serum medium more than control group, and experimental group G
0/ G
1phase, cell percentages was much larger than G
2/ M the phase, illustrate thus, stem cell serum-free culture medium provided by the invention can extend the G of cell
0/ G
1phase and the cultivation of the maintenance high cell densities of long period, under the hormesis of corresponding inductor, be conducive to mescenchymal stem cell and change into more object product.
In sum, stem cell serum-free culture medium provided by the invention, serum is replaced by adding recruitment factor in basic medium, the exogenous pollution that serum not only can be avoided to bring and the cytotoxic effect of serum component, and not clear serum component can be avoided the impact of cell cultures and experimental study; In addition, stem cell serum-free culture medium comparison of ingredients is clear and definite, uniform quality, is conducive to the stability improving cellular product production, improves the repeatability of cell cultures and test-results, and be easy to purifying cells product; In addition, serum-free culture provided by the invention also helps the G extending cell
0/ G
1phase, under the hormesis of cell induction agent, be conducive to differentiating more object product.
More than be described in conjunction with embodiments of the invention; 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 (9)
1. a stem cell serum-free culture medium, comprises basic medium, it is characterized in that: also comprise Regular Insulin, glutamine, the foodstuff additive of iron content, Sodium Selenite, progesterone and butanediamine.
2. stem cell serum-free culture medium according to claim 1, is characterized in that, the foodstuff additive of described iron content comprise in Transferrins,iron complexes, ironic citrate, ferric cacodylate and Gluconate Ferrecex one or more.
3. stem cell serum-free culture medium according to claim 1, it is characterized in that, the concentration that in described stem cell serum-free culture medium, the concentration of Regular Insulin is 1-15 μ g/ml, the concentration of glutamine is 1-5mmol/l, the concentration of the foodstuff additive of iron content is 20-200ng/ml, the concentration of Sodium Selenite is 20-50nmol/l, the concentration of progesterone is 5-30mmol/l and butanediamine is 10-100 μm of ol/l.
4. stem cell serum-free culture medium according to claim 1, is characterized in that, also comprises HEPES and penicillin.
5. stem cell serum-free culture medium according to claim 4, is characterized in that, the concentration of described HEPES is 2-20mmol/l, and the concentration of penicillin is 10-100mg/ml.
6. stem cell serum-free culture medium according to claim 1, is characterized in that, also comprises glucose, NaHCO
3, soybean pancreatin inhibitor, linolic acid and Streptomycin sulphate.
7. stem cell serum-free culture medium according to claim 6, is characterized in that, the massfraction of described glucose is 0.1-1%, NaHCO
3concentration be 1-10mmol/l, the massfraction of soybean pancreatin inhibitor is 0.05-0.2%, and linoleic concentration is 10-100 μm of ol/l, and the concentration of Streptomycin sulphate is 10-100mg/ml.
8. stem cell serum-free culture medium according to claim 1, is characterized in that, described basic medium is DMEM/F12 substratum.
9. a cultural method for stem cell, is characterized in that, comprises the steps:
Obtain stem cell, adopt the stem cell serum-free culture medium as described in any one of claim 1-8 to carry out vitro culture.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510518943.8A CN105087479A (en) | 2015-08-21 | 2015-08-21 | Stem cell serum-free culture medium and stem cell culture method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510518943.8A CN105087479A (en) | 2015-08-21 | 2015-08-21 | Stem cell serum-free culture medium and stem cell culture method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105087479A true CN105087479A (en) | 2015-11-25 |
Family
ID=54568815
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510518943.8A Pending CN105087479A (en) | 2015-08-21 | 2015-08-21 | Stem cell serum-free culture medium and stem cell culture method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105087479A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105907705A (en) * | 2016-06-28 | 2016-08-31 | 广州市搏克生物技术有限公司 | Multi-potent stem cell culture medium |
| CN106256900A (en) * | 2016-08-12 | 2016-12-28 | 浙江译美生物科技有限公司 | A kind of stem cell cultivating system of non-animal derived property |
| WO2019191402A1 (en) * | 2018-03-29 | 2019-10-03 | The University Of North Carolina At Chapel Hill | Stem/progenitor cells from duodenal brunner's glands and methods of isolating and using them |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN102946914A (en) * | 2010-04-12 | 2013-02-27 | 思百博技术股份公司 | Methods and combination |
| CN102994441A (en) * | 2012-09-19 | 2013-03-27 | 上海瀚康生物医药科技有限公司 | Cell culture medium, and preparation method and use thereof |
| WO2014118810A1 (en) * | 2013-02-01 | 2014-08-07 | Euroclone Spa | Serum-free culture medium for cancer stem cells |
-
2015
- 2015-08-21 CN CN201510518943.8A patent/CN105087479A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102946914A (en) * | 2010-04-12 | 2013-02-27 | 思百博技术股份公司 | Methods and combination |
| 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 |
| CN102994441A (en) * | 2012-09-19 | 2013-03-27 | 上海瀚康生物医药科技有限公司 | Cell culture medium, and preparation method and use thereof |
| WO2014118810A1 (en) * | 2013-02-01 | 2014-08-07 | Euroclone Spa | Serum-free culture medium for cancer stem cells |
Non-Patent Citations (2)
| Title |
|---|
| ZILONG LIU ET AL.: "Ethanol Suppresses PGC-1a Expression by Interfering with the cAMP-CREB Pathway in Neuronal Cells", 《PLOS ONE》 * |
| 陈锋: "无血清细胞培养基的主要补充因子及研究进展", 《海峡药学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105907705A (en) * | 2016-06-28 | 2016-08-31 | 广州市搏克生物技术有限公司 | Multi-potent stem cell culture medium |
| CN105907705B (en) * | 2016-06-28 | 2020-01-24 | 广州市搏克生物技术有限公司 | Pluripotent stem cell culture medium |
| CN106256900A (en) * | 2016-08-12 | 2016-12-28 | 浙江译美生物科技有限公司 | A kind of stem cell cultivating system of non-animal derived property |
| WO2019191402A1 (en) * | 2018-03-29 | 2019-10-03 | The University Of North Carolina At Chapel Hill | Stem/progenitor cells from duodenal brunner's glands and methods of isolating and using them |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105087481A (en) | Serum-free culture medium and stem cell culture method | |
| Eibes et al. | Maximizing the ex vivo expansion of human mesenchymal stem cells using a microcarrier-based stirred culture system | |
| CN103243070B (en) | Stem cell medium and application thereof | |
| CN101864393B (en) | Serum-free culture medium without animal origin components for culturing Vero cell micro-carrier | |
| CN102634482B (en) | Serum-free complete medium for mesenchymal stem cell | |
| CN102827810B (en) | Non-animal-source serum-free culture medium for umbilical cord blood stem cells | |
| KR20190112760A (en) | Multipotent Stem Cell Proliferation Accelerator | |
| CN104805054A (en) | Serum-free medium of stem cell | |
| CN104694470B (en) | A kind of stem cell serum-free culture medium | |
| CN105154386A (en) | Special culture medium and culture method for long-term maintenance, propagation, and subcultring of human hepatocyte | |
| CN105076114A (en) | Serum-free preservation liquid for umbilical cord mesenchymal stem cells and application of serum-free preservation liquid | |
| CN102311938A (en) | Serum-free medium for culturing hepatic cells | |
| CN105420179A (en) | Method for simultaneously extracting epithelial cells and mesenchymal stem cells from umbilical cord and placenta amnion tissues | |
| CN105039248B (en) | Tree shrew mesenchymal stem cell culture systems | |
| CN105087465A (en) | Hepatocyte serum-free culture medium | |
| CN106318906A (en) | Method for large-scale culture of human umbilical cord mesenchymal stem cells | |
| CN105087479A (en) | Stem cell serum-free culture medium and stem cell culture method | |
| CN104651305A (en) | Method for acquiring bioactive proteins by utilizing umbilical cord mesenchymal stem cells | |
| CN106929470A (en) | It is a kind of for derived mesenchymal stem cells in vitro culture and amplification serum free medium | |
| CN103320385B (en) | Humanized's differentiated hepatoma cell strain HL1017 and construction process thereof | |
| CN103421740B (en) | In-vitro culture and proliferation method for human mesenchymal stem cells | |
| CN106754657A (en) | A kind of serum free medium of monkey embryonic stem cell | |
| CN104726401A (en) | Method for improving success rate of umbilical cord blood mesenchymal stem cell culture by using placental mesenchymal stem cells | |
| CN104450610B (en) | Subculture method of human amniotic mesenchymal stem cells | |
| CN109430251A (en) | A kind of store method of the preservation liquid of adipose tissue and preparation method thereof with adipose tissue |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information |
Address after: 518057, room 2, building 10, Shenzhen biological incubation center, No. 302, Nanshan District hi tech, Shenzhen, Guangdong Applicant after: ISTEM REGENERATIVE MEDICINE SCI-TECH CO., LTD. Address before: 518057, Guangdong Province, Nanshan District high tech Zone, South Ring Road, No. 29, No. 02 building, international students, building No. 15 Applicant before: ISTEM REGENERATIVE MEDICINE SCI-TECH CO., LTD. |
|
| COR | Change of bibliographic data | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20151125 |
|
| WD01 | Invention patent application deemed withdrawn after publication |