Method for separation and proliferation of human urine derived mesenchymal stem cells
A stem cell and cell technology, applied in the field of biomedicine, can solve the problems of easy serum availability, high price, and high cost of serum acquisition, and achieve the effects of maintaining stem cell characteristics, vigorous cell proliferation, and reducing the risk of cell contamination.
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Embodiment 1
[0036] This embodiment provides a low-serum medium for isolating human urine-derived mesenchymal stem cells, including basal medium and additives; the basal medium is a mixture of DMEM medium, F12-K medium and KSFM medium system, the volume ratio of DMEM medium, F12-K medium and KSFM medium is 1:2:2; taking the volume of basal medium as a benchmark, the additives include: glutamine 0.2mM, antibiotic 10U / mL, Recombinant human insulin 1×10 -3 mg / mL, human epidermal growth factor 1×10 -6 mg / mL, platelet-derived growth factor 1×10 -6 mg / mL, basic fibroblast growth factor 1×10 -6 mg / mL, human transferrin 1×10 -3 mg / mL, selenous acid 1×10 -3 mg / mL, glycine 1mg / L, L-alanine 1mg / L, L-asparagine 1mg / L, L-aspartic acid 1mg / L, L-glutamic acid 1mg / L, L-proline Acid 1mg / L, L-serine 1mg / L, triiodothyronine 1×10 -6 mg / mL, fetal bovine serum 0.1ul / mL, hydrocortisone 0.1uM, epinephrine 1×10 -4 mg / mL.
[0037] Further, this embodiment provides a method for using the culture medium to is...
Embodiment 2
[0042] This embodiment provides a serum-free medium for isolating human urine-derived mesenchymal stem cells, including basal medium and additives; the basal medium is a mixture of DMEM medium, F12-K medium and KSFM medium System, the volume ratio of DMEM medium, F12-K medium and KSFM medium is 2:2:1; based on the volume of basal medium, the additives include: glutamine 2mM, antibiotic 100U / mL, recombinant Human insulin 0.1mg / mL, human epidermal growth factor 5×10 -5 mg / mL, platelet-derived growth factor 5×10 -5 mg / mL, basic fibroblast growth factor 5×10 -5 mg / mL, human transferrin 5×10 -2 mg / mL, selenous acid 8×10 -2 mg / mL, glycine 20mg / L, L-alanine 20mg / L, L-asparagine 20mg / L, L-aspartic acid 20mg / L, L-glutamic acid 20mg / L, L-proline Acid 20mg / L, L-serine 20mg / L, triiodothyronine 5×10 -5 mg / mL, adhesion factor 1×10 -2 mg / mL, albumin 50mg / mL, vitamin 1×10 -2 mg / mL, hydrocortisone 10uM, epinephrine 1×10 -2 mg / mL.
[0043] Further, this embodiment provides a method for...
Embodiment 3
[0048] This embodiment provides a low-serum medium for isolating human urine-derived mesenchymal stem cells, including basal medium and additives; the basal medium is a mixture of DMEM medium, F12-K medium and KSFM medium System, the volume ratio of DMEM medium, F12-K medium and KSFM medium is 2:1:2; Based on the volume of basal medium, the additives include: glutamine 1.1mM, antibiotic 55U / mL, Recombinant human insulin 4.55×10 -2 mg / mL, human epidermal growth factor 2.55×10 -5 mg / mL, platelet-derived growth factor 2.55×10 -5 mg / mL, basic fibroblast growth factor 2.55×10 -5 mg / mL, human transferrin 2.55×10 -2 mg / mL, selenous acid 3.95×10 -2 mg / mL, glycine 10.5mg / L, L-alanine 10.5mg / L, L-asparagine 10.5mg / L, L-aspartic acid 10.5mg / L, L-glutamic acid 10.5mg / L , L-proline 10.5mg / L, L-serine 10.5mg / L, triiodothyronine 2.55×10 -5 mg / mL, fetal bovine serum 25.75ul / mL, hydrocortisone 5.95uM, epinephrine 5.95×10 -3 mg / mL.
[0049] Further, this embodiment provides a method for...
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