CN109988746A - A kind of mescenchymal stem cell adipogenic induction differentiation method - Google Patents

A kind of mescenchymal stem cell adipogenic induction differentiation method Download PDF

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CN109988746A
CN109988746A CN201910037685.XA CN201910037685A CN109988746A CN 109988746 A CN109988746 A CN 109988746A CN 201910037685 A CN201910037685 A CN 201910037685A CN 109988746 A CN109988746 A CN 109988746A
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stem cell
mescenchymal stem
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adipogenic induction
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崔晓燕
徐卓
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Shanghai Baonian Biotechnology Co Ltd
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Abstract

The invention discloses a kind of mescenchymal stem cell adipogenic induction differentiation methods, comprising steps of 1) recovery mescenchymal stem cell is to growing to 70%-90%；2) mescenchymal stem cell is digested, complete medium culture 1-3 days is used；3) reach 70%-90% to cell fusion degree, siphon away complete medium, mescenchymal stem cell adipogenic induction differential medium is added (to be made of following components: DMEM/F12 basal medium, volumetric concentration 8%-12% fetal calf serum, 8-12 μ g/ml insulin, 1 ‰ penicillin, 1 ‰ streptomysins, 0.8-1.2 μm of ol/L dexamethasone, 100-500 μm of ol/L 3-isobutyl-1-methylxanthine) culture, culture medium was changed every 3-4 days；Successive induction, visual cell's upgrowth situation and at rouge situation depending on.The method of the present invention operation is appropriate, simplifies induction differentiation step, adipogenic induction differentiation rate is very high.

Description

A kind of mescenchymal stem cell adipogenic induction differentiation method
Technical field
The present invention relates to field of biotechnology, and in particular to a kind of mescenchymal stem cell adipogenic induction differentiation method.
Background technique
It is that there is height from the mescenchymal stem cell (mesenchymal stem cells, MSC) of foetal mesoderm The multipotential stem cell of self-renewal capacity and multi-lineage potential, be widely present in marrow, fat, amnion, placenta, Cord blood and It, can Induction of committed differentiation lipoblast, osteoblast, cardiac muscle cell, islet-like cells, intravascular in vitro in umbilical cord tissue Chrotoplast and epidermal cell etc..Except this, human mesenchymal stem cell have low immunogenicity, compared with low-risk pathogen infectious rate and Oncogenicity, clinical application are highly-safe.In addition, its materials is simple and convenient, no ethics limitation is protected in continuous passage culture and freezing After depositing, its biological characteristics is still kept.Human umbilical cord mesenchymal stem cells after transplanting can secrete various effectors, play immune Adjustment effect, it is clear to the curative effect of various damages, regression and diseases associated with inflammation, in stem-cell therapy, organizational project treatment and again It is of great significance in raw medical research application.
The Analytical Chemical Experiment of mescenchymal stem cell adipogenic induction from the angle of science demonstrate MSC have be directed differentiation to it is fatty thin The biological characteristics of born of the same parents are laid a good foundation for the basic research of stem cell.
Currently, at rouge induction cultural method general step are as follows:
1. preparing culture medium A liquid (basal medium A, fetal calf serum, mycillin, glutamine, insulin, 3- isobutyl Base -1- methyl xanthine, Rosiglitazone, dexamethasone) and culture medium B liquid (basal medium B, fetal calf serum, glutamine, Mycillin, insulin).
2. being digested after cell growth fusion rate to be checked reaches 80%-90% with 0.25% pancreatin.
3. by the cell digested by density 2 × 104A/cm2It is seeded in six orifice plates, every hole is added 2ml and cultivates completely Base.
4. changing liquid every three days, merged until cell confluency reaches 100% or crosses.
5. after carefully siphoning away complete medium, 2ml induced medium A liquid is added.
6. after induction three days, siphoning away A liquid, 2ml is added and maintains culture medium B liquid.After for 24 hours, B liquid is siphoned away, gains A induction.
7.A liquid and B liquid replace after 3-5 use (12-20 days), continue to maintain culture 4-7 days with B liquid, until fat granule becomes Enough to big, circle.B liquid maintenance period needed to renew every 2-3 days fresh B liquid.
Existing at rouge induction culture medium, that there are induced efficiencies is not high, undesirable at rouge effect, and cell is impure etc. asks Topic.
Patent CN104830757A, 2015.08.12 disclose a kind of mescenchymal stem cell adipogenic induction differential medium And preparation method thereof.The culture medium includes DMEM/F12 culture medium, is also comprised the following components and its concentration: FBS 5~50% Percent by volume, 0.5~5% percent by volume of glutamine, 0.5~5% percent by volume of antibiotic, Indomethacin 50~ 500 μM, 50~500nM of insulin, 5~50nM of dexamethasone, 0.5~5 μM of 3-isobutyl-1-methylxanthine and hydrochloric acid method are relaxed 0.05~0.5 μM of ground that.The mescenchymal stem cell is selected between mesenchymal stem cell, umbilical cord mesenchymal stem cells or amnion Mesenchymal stem cells.Mescenchymal stem cell adipogenic induction differentiation method includes the following steps: a) to cultivate mescenchymal stem cell；B) to thin When born of the same parents' convergence degree meets the requirements, the culture medium in the step a) is discarded, mescenchymal stem cell as described above is added and is lured at rouge Lead differential medium culture 2~4 weeks.The mescenchymal stem cell adipogenic induction differential medium that the invention provides has induced efficiency The advantages that high.
However, the inductive differentiation medium ingredient that the above mescenchymal stem cell adipogenic induction atomization uses is still more multiple It is miscellaneous, increase operation difficulty and cost.Have no the mescenchymal stem cell adipogenic induction differentiation method being more simple and efficient.
Summary of the invention
The present invention aiming at the shortcomings in the prior art, provides a kind of mescenchymal stem cell adipogenic induction being more simple and efficient Differentiation method.
A kind of mescenchymal stem cell adipogenic induction differential medium, the mescenchymal stem cell adipogenic induction point are provided first Change culture medium to be made of following components: DMEM/F12 basal medium, volumetric concentration 8%-12% fetal calf serum, 8-12 μ g/ml Insulin, 0.8 ‰ -1.2 ‰ penicillin of mass concentration, 0.8 ‰ -1.2 ‰ streptomysin of mass concentration are filled in 0.8-1.2 μm of ol/L Meter Song, 100-500 μm of ol/L 3-isobutyl-1-methylxanthines.
More preferably, the mescenchymal stem cell adipogenic induction differential medium is made of following components: the basis DMEM/F12 Culture medium, 10% fetal calf serum of volumetric concentration, 10 μ g/ml insulin, 1 ‰ penicillin of mass concentration, 1 ‰ strepto- of mass concentration Element, 1 μm of ol/L dexamethasone, 300 μm of ol/L 3-isobutyl-1-methylxanthines.
More preferably, the DMEM/F12 basic media components are as follows:
The mescenchymal stem cell adipogenic induction differential medium is also provided and is divided into fat in inducing mesenchymal stem cell Application in cell.
The mescenchymal stem cell is that fat mesenchymal stem cell, umbilical cord mesenchymal stem cells, medulla mesenchyma are dry thin Born of the same parents, amnion mesenchymal stem cell or placenta mesenchyma stem cell.
A kind of mescenchymal stem cell adipogenic induction differentiation method is also provided, comprising the following steps:
1) recovery mescenchymal stem cell grows to 70%-90% to mescenchymal stem cell；
2) cell is placed in 37 DEG C, 5%CO using complete medium by digestion mescenchymal stem cell2Incubator in cultivate 1-3 days；
3) reach 70%-90% to cell fusion degree, siphon away complete medium, mescenchymal stem cell as described above is added Cell is placed in 37 DEG C, 5%CO by adipogenic induction differential medium2Incubator in cultivate, renewed every 3-4 days fresh as above The mescenchymal stem cell adipogenic induction differential medium；Successive induction, visual cell's upgrowth situation and at rouge situation depending on.
More preferably, the step 3) successive induction is specially successive induction 3 weeks or more.
More preferably, the complete medium is DMEM complete medium.
The invention has the benefit that
Culture medium agents useful for same component of the invention is few, and ingredient is simpler, and abductive approach of the invention has only used one kind Culture medium, operation is appropriate, generally simplifies mescenchymal stem cell to the induction of fat cell and breaks up operation.But through oil red O Dyeing is it has been observed that cell adipogenic induction differentiation rate of the present invention is very high under identical induction time.
Detailed description of the invention
Fig. 1: the oil red O stain result of embodiment 11.
Specific embodiment
Present inventor is based on mescenchymal stem cell adipogenic induction abundant and breaks up experience, known cell characteristics, culture Association between influence of each ingredient to cell induction differentiation in base, especially the difference factors, mescenchymal stem cell is lured at rouge It leads differential medium and method is improved, the present invention is completed based on this.
Specific embodiment provided by the invention is further described below.
Material source in following embodiment:
Umbilical cord mesenchymal stem cells, fat mesenchymal stem cell are purchased from ATCC.
DMEM/F12 basal medium, DMED low sugar complete medium, fetal calf serum, insulin are purchased from Gibco；Or for certainly System is formulated as follows:
Penicillin, streptomysin, dexamethasone, 3-isobutyl-1-methylxanthine are purchased from Chinese medicines group.
Embodiment 1
1. preparation umbilical cord mesenchymal stem cells adipogenic induction differential medium (self-control DMEM/F12 basal medium, 10% Fetal calf serum, 10 μ g/ml insulin, 1 ‰ penicillin, 1 ‰ streptomysins, 1 μm of ol/L dexamethasone, 300 μm of ol/L 3- isobutyls Base -1- methyl xanthine).
2. after cell recovery, cell to be detected grows to 80% or so, passed on.Generally do 3 holes in six orifice plates, one Control, an experimental group, a multiple holes are not induced.
3. by the mescenchymal stem cell digested according to 1 × 104cells/cm2Cell density be seeded in six orifice plates, 3ml DMEM complete medium is added in every hole, cell is placed in 37 DEG C, 5%CO2Incubator in cultivated.
4. about 2 days, cell fusion degree reached 80% or so, carefully mescenchymal stem cell complete medium is siphoned away, to 3ml mescenchymal stem cell adipogenic induction differential medium is added in the six experimental group holes of orifice plate two.
5. renewing fresh mescenchymal stem cell adipogenic induction differential medium every 3 or 4 days.
6. successive induction 3 weeks or more, visual cell's upgrowth situation and at rouge situation depending on.
Embodiment 2
1. preparing umbilical cord mesenchymal stem cells adipogenic induction differential medium (self-control DMEM/F12 basal medium, 8% tire Cow's serum, 8 μ g/ml insulin, 0.8 ‰ penicillin, 1 ‰ streptomysins, 1.2 μm of ol/L dexamethasone, 500 μm of ol/L 3- isobutyls Base -1- methyl xanthine).
2. after cell recovery, cell to be detected grows to 90% or so, passed on.Generally do 3 holes in six orifice plates, one Control, an experimental group, a multiple holes are not induced.
3. by the mescenchymal stem cell digested according to 1 × 104cells/cm2Cell density be seeded in six orifice plates, 3ml DMEM complete medium is added in every hole, cell is placed in 37 DEG C, 5%CO2Incubator in cultivated.
4. about 1 day, cell fusion degree reached 70% or so, carefully mescenchymal stem cell complete medium is siphoned away, to 3ml mescenchymal stem cell adipogenic induction differential medium is added in the six experimental group holes of orifice plate two.
5. renewing fresh mescenchymal stem cell adipogenic induction differential medium every 3 or 4 days.
6. successive induction 3 weeks or more, visual cell's upgrowth situation and at rouge situation depending on.
Embodiment 3
1. preparation umbilical cord mesenchymal stem cells adipogenic induction differential medium (self-control DMEM/F12 basal medium, 10% Fetal calf serum, 12 μ g/ml insulin, 0.8 ‰ penicillin, 1.2 ‰ streptomysins, 0.8 μm of ol/L dexamethasone, 100 μm of ol/L 3- Isobutyl group -1- methyl xanthine).
2. after cell recovery, cell to be detected grows to 80% or so, passed on.Generally do 3 holes in six orifice plates, one Control, an experimental group, a multiple holes are not induced.
3. by the mescenchymal stem cell digested according to 1 × 104cells/cm2Cell density be seeded in six orifice plates, 3ml DMED complete medium is added in every hole, cell is placed in 37 DEG C, 5%CO2Incubator in cultivated.
4. about 3 days, cell fusion degree reached 80% or so, carefully mescenchymal stem cell complete medium is siphoned away, to 3ml mescenchymal stem cell adipogenic induction differential medium is added in the six experimental group holes of orifice plate two.
5. renewing fresh mescenchymal stem cell adipogenic induction differential medium every 3 or 4 days.
6. successive induction 3 weeks or more, visual cell's upgrowth situation and at rouge situation depending on.
Embodiment 4
1. preparation umbilical cord mesenchymal stem cells adipogenic induction differential medium (Gibco DMEM/F12 basal medium, 8% Fetal calf serum, 10 μ g/ml insulin, 1 ‰ penicillin, 1 ‰ streptomysins, 1.2 μm of ol/L dexamethasone, 200 μm of ol/L 3- isobutyls Base -1- methyl xanthine).
2. after cell recovery, cell to be detected grows to 90% or so, passed on.Generally do 3 holes in six orifice plates, one Control, an experimental group, a multiple holes are not induced.
3. by the mescenchymal stem cell digested according to 1 × 104cells/cm2Cell density be seeded in six orifice plates, 3ml DMED complete medium is added in every hole, cell is placed in 37 DEG C, 5%CO2Incubator in cultivated.
4. about 2 days, cell fusion degree reached 80% or so, carefully mescenchymal stem cell complete medium is siphoned away, to 3ml mescenchymal stem cell adipogenic induction differential medium is added in the six experimental group holes of orifice plate two.
5. renewing fresh mescenchymal stem cell adipogenic induction differential medium every 3 or 4 days.
6. successive induction 3 weeks or more, visual cell's upgrowth situation and at rouge situation depending on.
Embodiment 5
1. preparation umbilical cord mesenchymal stem cells adipogenic induction differential medium (Gibco DMEM/F12 basal medium, 10% fetal calf serum, 10 μ g/ml insulin, 1.2 ‰ penicillin, 0.8 ‰ streptomysins, 1.2 μm of ol/L dexamethasone, 400 μm of ol/ L 3-isobutyl-1-methylxanthine).
2. after cell recovery, cell to be detected grows to 70% or so, passed on.Generally do 3 holes in six orifice plates, one Control, an experimental group, a multiple holes are not induced.
3. by the mescenchymal stem cell digested according to 1 × 104cells/cm2Cell density be seeded in six orifice plates, 3ml DMED complete medium is added in every hole, cell is placed in 37 DEG C, 5%CO2Incubator in cultivated.
4. about 3 days, cell fusion degree reached 90% or so, carefully mescenchymal stem cell complete medium is siphoned away, to 3ml mescenchymal stem cell adipogenic induction differential medium is added in the six experimental group holes of orifice plate two.
5. renewing fresh mescenchymal stem cell adipogenic induction differential medium every 3 or 4 days.
6. successive induction 3 weeks or more, visual cell's upgrowth situation and at rouge situation depending on.
Embodiment 6
1. preparation fat mesenchymal stem cell adipogenic induction differential medium (self-control DMEM/F12 basal medium, 10% Fetal calf serum, 10 μ g/ml insulin, 1 ‰ penicillin, 1 ‰ streptomysins, 1 μm of ol/L dexamethasone, 300 μm of ol/L 3- isobutyls Base -1- methyl xanthine).
2. after cell recovery, cell to be detected grows to 80% or so, passed on.Generally do 3 holes in six orifice plates, one Control, an experimental group, a multiple holes are not induced.
3. by the mescenchymal stem cell digested according to 1 × 104cells/cm2Cell density be seeded in six orifice plates, 3ml DMED complete medium is added in every hole, cell is placed in 37 DEG C, 5%CO2Incubator in cultivated.
4. about 1.5 days, cell fusion degree reached 80% or so, carefully mescenchymal stem cell complete medium is siphoned away, 3ml mescenchymal stem cell adipogenic induction differential medium is added into six orifice plates, two experimental group holes.
5. renewing fresh mescenchymal stem cell adipogenic induction differential medium every 3 or 4 days.
6. successive induction 3 weeks or more, visual cell's upgrowth situation and at rouge situation depending on.
Embodiment 7
1. preparing fat mesenchymal stem cell adipogenic induction differential medium (self-control DMEM/F12 basal medium, 8% tire Cow's serum, 8 μ g/ml insulin, 1 ‰ penicillin, 1 ‰ streptomysins, 1.2 μm of ol/L dexamethasone, 500 μm of ol/L 3- isobutyl groups- 1- methyl xanthine).
2. after cell recovery, cell to be detected grows to 90% or so, passed on.Generally do 3 holes in six orifice plates, one Control, an experimental group, a multiple holes are not induced.
3. by the mescenchymal stem cell digested according to 1 × 104cells/cm2Cell density be seeded in six orifice plates, 3ml DMED complete medium is added in every hole, cell is placed in 37 DEG C, 5%CO2Incubator in cultivated.
4. about 1.5 days, cell fusion degree reached 80% or so, carefully mescenchymal stem cell complete medium is siphoned away, 3ml mescenchymal stem cell adipogenic induction differential medium is added into six orifice plates, two experimental group holes.
5. renewing fresh mescenchymal stem cell adipogenic induction differential medium every 3 or 4 days.
6. successive induction 3 weeks or more, visual cell's upgrowth situation and at rouge situation depending on.
Embodiment 8
1. preparation fat mesenchymal stem cell adipogenic induction differential medium (Gibco DMEM/F12 basal medium, 10% fetal calf serum, 12 μ g/ml insulin, 1 ‰ penicillin, 1 ‰ streptomysins, 0.8 μm of ol/L dexamethasone, 100 μm of ol/L 3- Isobutyl group -1- methyl xanthine).
2. after cell recovery, cell to be detected grows to 80% or so, passed on.Generally do 3 holes in six orifice plates, one Control, an experimental group, a multiple holes are not induced.
3. by the mescenchymal stem cell digested according to 1 × 104cells/cm2Cell density be seeded in six orifice plates, 3ml DMED complete medium is added in every hole, cell is placed in 37 DEG C, 5%CO2Incubator in cultivated.
4. about 2 days, cell fusion degree reached 80% or so, carefully mescenchymal stem cell complete medium is siphoned away, to 3ml mescenchymal stem cell adipogenic induction differential medium is added in the six experimental group holes of orifice plate two.
5. renewing fresh mescenchymal stem cell adipogenic induction differential medium every 3 or 4 days.
6. successive induction 3 weeks or more, visual cell's upgrowth situation and at rouge situation depending on.
Embodiment 9
1. preparation navel fat mesenchymal stem cell adipogenic induction differential medium (self-control DMEM/F12 basal medium, 8% Fetal calf serum, 10 μ g/ml insulin, 1 ‰ penicillin, 0.8 ‰ streptomysins, 1.2 μm of ol/L dexamethasone, 200 μm of ol/L 3- are different Butyl -1- methyl xanthine).
2. after cell recovery, cell to be detected grows to 90% or so, passed on.Generally do 3 holes in six orifice plates, one Control, an experimental group, a multiple holes are not induced.
3. by the mescenchymal stem cell digested according to 1 × 104cells/cm2Cell density be seeded in six orifice plates, 3ml DMED complete medium is added in every hole, cell is placed in 37 DEG C, 5%CO2Incubator in cultivated.
4. about 1.5 days, cell fusion degree reached 80% or so, carefully mescenchymal stem cell complete medium is siphoned away, 3ml mescenchymal stem cell adipogenic induction differential medium is added into six orifice plates, two experimental group holes.
5. renewing fresh mescenchymal stem cell adipogenic induction differential medium every 3 or 4 days.
6. successive induction 3 weeks or more, visual cell's upgrowth situation and at rouge situation depending on.
Embodiment 10
1. preparation fat mesenchymal stem cell adipogenic induction differential medium (self-control DMEM/F12 basal medium, 10% Fetal calf serum, 10 μ g/ml insulin, 1.2 ‰ penicillin, 1 ‰ streptomysins, 1.2 μm of ol/L dexamethasone, 400 μm of ol/L 3- are different Butyl -1- methyl xanthine).
2. after cell recovery, cell to be detected grows to 70% or so, passed on.Generally do 3 holes in six orifice plates, one Control, an experimental group, a multiple holes are not induced.
3. by the mescenchymal stem cell digested according to 1 × 104cells/cm2Cell density be seeded in six orifice plates, 3ml DMED complete medium is added in every hole, cell is placed in 37 DEG C, 5%CO2Incubator in cultivated.
4. about 2.5 days, cell fusion degree reached 90% or so, carefully mescenchymal stem cell complete medium is siphoned away, 3ml mescenchymal stem cell adipogenic induction differential medium is added into six orifice plates, two experimental group holes.
5. renewing fresh mescenchymal stem cell adipogenic induction differential medium every 3 or 4 days.
6. successive induction 3 weeks or more, visual cell's upgrowth situation and at rouge situation depending on.
Embodiment 11
By mescenchymal stem cell adipogenic induction differentiation method of the present invention and use Applied CellTMHuman mesenchymal is dry thin Born of the same parents' induction is compared at the method for rouge differentiation agents box.Equal successive induction 3 weeks, then carry out oil red O stain.As shown in Figure 1, A is using kit inducing umbilical cord mesenchymal stem as a result, B is the induction of 1 method of embodiment as a result, C is 6 method of embodiment The result of induction.It can be seen that identical induction time, the visible a large amount of fat drips of the method for the present invention are formed.By plane optical densitometric method to each group Fat drips formational situation carries out quantitative analysis, obtains example 1 group and 6 groups of embodiment of fat drips quantity is kit group fat drips respectively 2.8 times of quantity and 2.5 times, example 1 group and 6 groups of embodiment of fat drips area are the 3.3 of kit group fat drips area respectively Times and 3.9 times.
Embodiment 12
Mescenchymal stem cell adipogenic induction differentiation method of the present invention is compared with the method for patent CN104830757A.
It is specific as described in Example 1 using the method inducing umbilical cord mesenchymal stem of the embodiment of the present invention 1.Induction training It supports 3 weeks.
It is specific as described in Example 6 using the method induced lipolysis mescenchymal stem cell of the embodiment of the present invention 6.Induction training It supports 3 weeks.
It utilizes the culture medium inducing umbilical cord mesenchymal stem of patent CN104830757A embodiment 1: taking P3 between umbilical cord Mesenchymal stem cells, with 5000/cm2Density be inoculated in six orifice plates, and with containing 10% (v/v) FBS DMEM/F12 cultivate Base is cultivated, and when cell confluency degree is up to 80%, is discarded culture medium, is separately added into 1 (patent of induced medium The culture medium of CN104830757A embodiment 1), it changes the liquid once within every 3 days.Fiber differentiation 3 weeks.
Utilize the culture medium induced lipolysis mescenchymal stem cell of patent CN104830757A embodiment 3:
1. after cell recovery, cell to be detected grows to 80% or so, passed on.Generally do 3 holes in six orifice plates, one Control, an experimental group, a multiple holes are not induced.
2. by the mescenchymal stem cell digested according to 1 × 104cells/cm2Cell density be seeded in six orifice plates, 3ml complete medium is added in every hole, cell is placed in 37 DEG C, 5%CO2Incubator in cultivated.
3. about 1.5 days, cell fusion degree reached 80% or so, carefully mescenchymal stem cell complete medium is siphoned away, The induced medium of 3ml patent CN104830757A embodiment 3 is added into six orifice plates, two experimental group holes.
4. renewing fresh induced medium every 3 or 4 days.
5. successive induction 3 weeks.
Induction 3 weeks, then carries out oil red O stain, is quantified by plane optical densitometric method to each group fat drips formational situation Analysis, the fat drips quantity for being calculated 1 group of the embodiment of the present invention is patent CN104830757A example 1 group fat drips quantity 2.6 times, fat drips area is its 3.1 times；6 groups of the embodiment of the present invention of fat drips quantity is 3 groups of patent CN104830757A embodiment 2.9 times of fat drips quantity, fat drips area are its 3.3 times.
It should be understood that above embodiments are only illustrative of the invention and is not intended to limit the scope of the invention.In addition, it should also be understood that, After reading the content taught by the present invention, those skilled in the art can make various modifications or changes to the present invention, these Equivalent form is also fallen within the scope of the appended claims of the present application.

Claims (8)

1. a kind of mescenchymal stem cell adipogenic induction differential medium, which is characterized in that be made of following components: DMEM/F12 base Basal culture medium, volumetric concentration 8%-12% fetal calf serum, 8-12 μ g/ml insulin, 0.8 ‰ -1.2 ‰ penicillin of mass concentration, 0.8 ‰ -1.2 ‰ streptomysin of mass concentration, 0.8-1.2 μm of ol/L dexamethasone, 100-500 μm of ol/L 3- isobutyl group -1- methyl Xanthine.
2. mescenchymal stem cell adipogenic induction differential medium according to claim 1, which is characterized in that by following components It constitutes: DMEM/F12 basal medium, 10% fetal calf serum of volumetric concentration, 10 μ g/ml insulin, 1 ‰ penicillin of mass concentration, 1 ‰ streptomysin of mass concentration, 1 μm of ol/L dexamethasone, 300 μm of ol/L 3-isobutyl-1-methylxanthines.
3. mescenchymal stem cell adipogenic induction differential medium according to claim 1 or 2, which is characterized in that described DMEM/F12 basic media components are as follows:
4. mescenchymal stem cell adipogenic induction differential medium of any of claims 1 or 2 breaks up in inducing mesenchymal stem cell For the application in fat cell.
5. application according to claim 4, which is characterized in that the mescenchymal stem cell is that fat mesenchymal is dry thin Born of the same parents, umbilical cord mesenchymal stem cells, mesenchymal stem cell, amnion mesenchymal stem cell or placenta mesenchyma stem cell.
6. a kind of mescenchymal stem cell adipogenic induction differentiation method, which comprises the following steps:
1) recovery mescenchymal stem cell grows to 70%-90% to mescenchymal stem cell；
2) cell is placed in 37 DEG C, 5%CO using complete medium by digestion mescenchymal stem cell2Incubator in cultivate 1-3 It；
3) reach 70%-90% to cell fusion degree, siphon away complete medium, it is dry that mesenchyma of any of claims 1 or 2 is added Cell is placed in 37 DEG C, 5%CO by cell adipogenic induction differential medium2Incubator in cultivate, renewed every 3-4 days fresh Mescenchymal stem cell adipogenic induction differential medium of any of claims 1 or 2；Successive induction, visual cell's upgrowth situation and at Depending on rouge situation.
7. mescenchymal stem cell adipogenic induction differentiation method according to claim 6, which is characterized in that step 3) is described to hold Continuous induction is specially successive induction 3 weeks or more.
8. mescenchymal stem cell adipogenic induction differentiation method according to claim 6, which is characterized in that the complete culture Base is DMEM complete medium.
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CN110257324A (en) * 2019-07-24 2019-09-20 安徽科门生物科技有限公司 A kind of culture medium and abductive approach for fat mesenchymal stem cell adipogenic induction
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CN112941017A (en) * 2019-12-10 2021-06-11 东莞市恩联干细胞生物科技研究院 Culture medium for inducing human mesenchymal stem cells to form fat and differentiate and preparation method thereof
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CN113980893A (en) * 2021-11-29 2022-01-28 广东普罗凯融生物医药科技有限公司 Culture medium for inducing dental pulp stem cells to differentiate into adipogenic cells, kit containing culture medium and application of culture medium
CN113980893B (en) * 2021-11-29 2023-03-24 广东普罗凯融生物医药科技有限公司 Culture medium for inducing dental pulp stem cells to differentiate into adipogenic cells, kit containing culture medium and application of culture medium

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