An Intermediate Concentration of Calcium with Antioxidant Supplement in Culture Medium Enhances Proliferation and Decreases the Aging of Bone Marrow Mesenchymal Stem Cells

Int J Mol Sci. 2021 Feb 20;22(4):2095. doi: 10.3390/ijms22042095.

Abstract

Human bone marrow stem cells (HBMSCs) are isolated from the bone marrow. Stem cells can self-renew and differentiate into various types of cells. They are able to regenerate kinds of tissue that are potentially used for tissue engineering. To maintain and expand these cells under culture conditions is difficult-they are easily triggered for differentiation or death. In this study, we describe a new culture formula to culture isolated HBMSCs. This new formula was modified from NCDB 153, a medium with low calcium, supplied with 5% FBS, extra growth factor added to it, and supplemented with N-acetyl-L-cysteine and L-ascorbic acid-2-phosphate to maintain the cells in a steady stage. The cells retain these characteristics as primarily isolated HBMSCs. Moreover, our new formula keeps HBMSCs with high proliferation rate and multiple linage differentiation ability, such as osteoblastogenesis, chondrogenesis, and adipogenesis. It also retains HBMSCs with stable chromosome, DNA, telomere length, and telomerase activity, even after long-term culture. Senescence can be minimized under this new formulation and carcinogenesis of stem cells can also be prevented. These modifications greatly enhance the survival rate, growth rate, and basal characteristics of isolated HBMSCs, which will be very helpful in stem cell research.

Keywords: cell aging; cell differentiation; cell proliferation; culture medium; senescence.

MeSH terms

  • Antigens, CD / metabolism
  • Antioxidants / pharmacology*
  • Biomarkers / metabolism
  • Calcium / pharmacology*
  • Cell Differentiation / drug effects
  • Cell Lineage / drug effects
  • Cell Proliferation / drug effects
  • Cell Separation
  • Cell Shape / drug effects
  • Cells, Cultured
  • Cellular Senescence* / drug effects
  • Culture Media / chemistry*
  • DNA Damage
  • Humans
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / metabolism
  • Osteoblasts / cytology
  • Osteoblasts / drug effects
  • Osteoblasts / metabolism
  • Telomerase / metabolism
  • Telomere Homeostasis
  • Tumor Suppressor p53-Binding Protein 1 / metabolism

Substances

  • Antigens, CD
  • Antioxidants
  • Biomarkers
  • Culture Media
  • TP53BP1 protein, human
  • Tumor Suppressor p53-Binding Protein 1
  • Telomerase
  • Calcium