A method for promoting directed differentiation of mesenchymal stem cells

Abstract

The invention relates to the field of stem cells, and provides a method for promoting directional differentiation of mesenchymal stem cells. The method for promoting directional differentiation of themesenchymal stem cells is used for solving the problem of low efficiency of differentiation of stem cells into bone cells. The method for promoting directional differentiation of the mesenchymal stemcells comprises the following steps: culturing the mesenchymal stem cells by adopting a DMEM culture medium, conducting adherent culture in a six-hole board after the cells thaw, conducting digestionby means of a 0.25% pancreatic enzyme every three days, and conducting passage; and inoculating a differentiation culture medium against cultured mesenchymal stem cells, and conducting induced differentiation culture, wherein a differentiation culture medium comprises insulin with a concentration of 3-5 <mu>g / ml, transferrin with a concentration of 0.05-0.08 <mu>g / ml, a growth hormone with a concentration of 1-3 ng / ml, glutathione with a concentration of 80-100 <mu>g / ml, L-glutamine with a concentration of 0.5-2 mM, 2-mercaptoethanol with a concentration of 20-50 <mu>M, sodium pyruvate with aconcentration of 0.5-0.7 mM, nonessential amino acids with a concentration of 0.05-0.1 mM, nanometer ferric oxide with a concentration of 8-12 <mu>M and the balance a DMEM culture medium. No biological serum is added into the culture mediums, and the nanometer oxide is introduced at the same time so that differentiation of the mesenchymal stem cells into bone cells can be significantly promoted.

Description

technical field

[0001] The invention relates to the field of stem cells, in particular to a method for promoting directed differentiation of mesenchymal stem cells. Background technique

[0002] Stem cells are a kind of cell type with self-renewal ability and multi-directional differentiation potential. It is the first cell in the process of human newborn cells from proliferation, migration, differentiation to maturity. The study of stem cells has become one of the most challenging and attractive fields in biology. Stem cells can be divided into embryonic stem cells and adult stem cells according to their source. Mesenchymal stem cells (MSCs, mesenehymal stem cells) are one of the adult stem cells. As a type of adult stem cells with self-renewal ability, mesenchymal stem cells can differentiate into various cell types. It can not only differentiate into a variety of mesenchymal-derived cell lineages, such as adipocytes, osteoblasts, chondrocytes, myocytes, etc., but also ...

Examples

Embodiment 1

[0021] A method for promoting the directional differentiation of mesenchymal stem cells, comprising: using DMEM medium to culture mesenchymal stem cells, culturing the cells in a 6-well plate after recovery, digesting with 0.25% trypsin every 2 days and subculture; The obtained mesenchymal stem cells were inoculated on the differentiation medium to induce differentiation culture; the differentiation medium included insulin 4 μg / ml, transferrin 0.06 μg / ml, growth hormone 2ng / ml, glutathione 90 μg / ml, L-glutamine 1.5mM, β-mercaptoethanol 25μM, sodium pyruvate 0.6mM, non-essential amino acid 0.08mM, nano-ferric oxide 10μM, and the balance was DMEM medium. The nanometer ferric oxide is modified nanometer ferric oxide. The preparation method of the modified ferric oxide is as follows: take 21 mass parts of ferric nitrate, 2 mass parts of silver nitrate, 17 mass parts of sodium hydroxide, and 4 mass parts of carbon nanotubes; dissolve ferric nitrate in 40 times the amount of anhydro...

Embodiment 2

[0024] A method for promoting the directional differentiation of mesenchymal stem cells, comprising: using DMEM medium to culture mesenchymal stem cells, culturing the cells in a 6-well plate after recovery, digesting with 0.25% trypsin every 2 days and subculture; The obtained mesenchymal stem cells were inoculated on the differentiation medium for induced differentiation culture; the differentiation medium included: insulin 3 μg / ml, transferrin 0.05 μg / ml, growth hormone 1 ng / ml, glutathione 80 μg / ml , L-glutamine 0.5mM, β-mercaptoethanol 20μM, sodium pyruvate 0.5mM, non-essential amino acid 0.05mM, nanometer ferric oxide 8μM, and the balance is DMEM medium. The nanometer ferric oxide is modified nanometer ferric oxide. The preparation method of the modified ferric oxide is as follows: take 20 parts by mass of ferric nitrate, 1 part by mass of silver nitrate, 15 parts by mass of sodium hydroxide, and 3 parts by mass of carbon nanotubes; dissolve ferric nitrate in 40 times th...

Embodiment 3

[0026] A method for promoting the directional differentiation of mesenchymal stem cells, comprising: using DMEM medium to culture mesenchymal stem cells, culturing the cells in a 6-well plate after recovery, digesting with 0.25% trypsin every 2 days and subculture; The obtained mesenchymal stem cells were inoculated on the differentiation medium for induced differentiation culture; the differentiation medium included: insulin 5 μg / ml, transferrin 0.08 μg / ml, growth hormone 3 ng / ml, glutathione 100 μg / ml , L-glutamine 2mM, β-mercaptoethanol 50μM, sodium pyruvate 0.7mM, non-essential amino acid 0.1mM, nano-ferric oxide 12μM, and the balance was DMEM medium. The nanometer ferric oxide is modified nanometer ferric oxide. The preparation method of the modified ferric oxide is as follows: take 25 parts by mass of ferric nitrate, 3 parts by mass of silver nitrate, 20 parts by mass of sodium hydroxide, and 5 parts by mass of carbon nanotubes; dissolve ferric nitrate in 40 times the am...

technical field

[0001] The invention relates to the field of stem cells, in particular to a method for promoting directed differentiation of mesenchymal stem cells. Background technique

[0002] Stem cells are a kind of cell type with self-renewal ability and multi-directional differentiation potential. It is the first cell in the process of human newborn cells from proliferation, migration, differentiation to maturity. The study of stem cells has become one of the most challenging and attractive fields in biology. Stem cells can be divided into embryonic stem cells and adult stem cells according to their source. Mesenchymal stem cells (MSCs, mesenehymal stem cells) are one of the adult stem cells. As a type of adult stem cells with self-renewal ability, mesenchymal stem cells can differentiate into various cell types. It can not only differentiate into a variety of mesenchymal-derived cell lineages, such as adipocytes, osteoblasts, chondrocytes, myocytes, etc., but also ...

Examples

Embodiment 1

[0021] A method for promoting the directional differentiation of mesenchymal stem cells, comprising: using DMEM medium to culture mesenchymal stem cells, culturing the cells in a 6-well plate after recovery, digesting with 0.25% trypsin every 2 days and subculture; The obtained mesenchymal stem cells were inoculated on the differentiation medium to induce differentiation culture; the differentiation medium included insulin 4 μg / ml, transferrin 0.06 μg / ml, growth hormone 2ng / ml, glutathione 90 μg / ml, L-glutamine 1.5mM, β-mercaptoethanol 25μM, sodium pyruvate 0.6mM, non-essential amino acid 0.08mM, nano-ferric oxide 10μM, and the balance was DMEM medium. The nanometer ferric oxide is modified nanometer ferric oxide. The preparation method of the modified ferric oxide is as follows: take 21 mass parts of ferric nitrate, 2 mass parts of silver nitrate, 17 mass parts of sodium hydroxide, and 4 mass parts of carbon nanotubes; dissolve ferric nitrate in 40 times the amount of anhydro...

Embodiment 2

[0024] A method for promoting the directional differentiation of mesenchymal stem cells, comprising: using DMEM medium to culture mesenchymal stem cells, culturing the cells in a 6-well plate after recovery, digesting with 0.25% trypsin every 2 days and subculture; The obtained mesenchymal stem cells were inoculated on the differentiation medium for induced differentiation culture; the differentiation medium included: insulin 3 μg / ml, transferrin 0.05 μg / ml, growth hormone 1 ng / ml, glutathione 80 μg / ml , L-glutamine 0.5mM, β-mercaptoethanol 20μM, sodium pyruvate 0.5mM, non-essential amino acid 0.05mM, nanometer ferric oxide 8μM, and the balance is DMEM medium. The nanometer ferric oxide is modified nanometer ferric oxide. The preparation method of the modified ferric oxide is as follows: take 20 parts by mass of ferric nitrate, 1 part by mass of silver nitrate, 15 parts by mass of sodium hydroxide, and 3 parts by mass of carbon nanotubes; dissolve ferric nitrate in 40 times th...

Embodiment 3

[0026] A method for promoting the directional differentiation of mesenchymal stem cells, comprising: using DMEM medium to culture mesenchymal stem cells, culturing the cells in a 6-well plate after recovery, digesting with 0.25% trypsin every 2 days and subculture; The obtained mesenchymal stem cells were inoculated on the differentiation medium for induced differentiation culture; the differentiation medium included: insulin 5 μg / ml, transferrin 0.08 μg / ml, growth hormone 3 ng / ml, glutathione 100 μg / ml , L-glutamine 2mM, β-mercaptoethanol 50μM, sodium pyruvate 0.7mM, non-essential amino acid 0.1mM, nano-ferric oxide 12μM, and the balance was DMEM medium. The nanometer ferric oxide is modified nanometer ferric oxide. The preparation method of the modified ferric oxide is as follows: take 25 parts by mass of ferric nitrate, 3 parts by mass of silver nitrate, 20 parts by mass of sodium hydroxide, and 5 parts by mass of carbon nanotubes; dissolve ferric nitrate in 40 times the am...

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