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

Chung-Da Yang,Hui-Ting Chen,Cheng-Jung Ho,Hsuan-Ti Huang,Je-Ken Chang,Lin Kang,Chung-Hwan Chen,Sung-Yen Lin,Mei-Ling Ho,Tsung-Lin Cheng,Yi-Shan Lin,Mon-Juan Lee,Shu-Chun Chuang

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.

Highlights

Mesenchymal cells (MSCs) are adult multipotent stem cells that can be isolated from various tissues such as bone marrow, adipose tissue, fetal membranes of term placenta, dental pulp, and ovarian follicular liquid
Dulbecco modified Eagle medium (DMEM) were about 2.2–2.7 folds in the late stage (e.g., 7th–8th passage) and the morphology is flat. These results show that Human bone marrow stem cells (HBMSCs) can proliferate more cell morphology is flat. These results show that HBMSC can proliferate passages and cells in BK medium than in DMEM medium
We report a new formula for isolating and culturing HBMSCs that belong to adult stem cells and are rare in the bone marrow [26]

Summary

Introduction

Mesenchymal cells (MSCs) are adult multipotent stem cells that can be isolated from various tissues such as bone marrow, adipose tissue, fetal membranes of term placenta, dental pulp, and ovarian follicular liquid. A soft blood-forming living tissue, is composed of bone cavities that contain fat, immature and mature blood cells, hematopoietic stem cells, and non-hematopoietic stem cells. Human bone mesenchymal stem cells (HBMSCs) belong to adult stem cells, like all stem cells, sharing at least two characteristics. They can make identical copies of themselves for long periods of time; this ability to proliferate is referred to as longterm self-renewal. They can give rise to mature cell types that have characteristic morphologies (shapes) and specialized functions [7,8]

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