Enhanced proliferation of human skeletal muscle precursor cells derived from elderly donors cultured in estimated physiological (5%) oxygen |
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Authors: | Sheree D Martin Fiona M Collier Mark A Kirkland Ken Walder Nicole Stupka |
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Institution: | (1) Institute for Technology, Research and Innovation, Deakin University, Pigdons Road, Waurn Ponds, VIC, 3217, Australia;(2) Barwon Biomedical Research, Barwon Health, Ryrie Street, Geelong, VIC, 3220, Australia;(3) School of Medicine, Deakin University, Pigdons Road, Waurn Ponds, VIC, 3217, Australia; |
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Abstract: | Human skeletal muscle precursor cells (myoblasts) have significant therapeutic potential and are a valuable research tool
to study muscle cell biology. Oxygen is a critical factor in the successful culture of myoblasts with low (1–6%) oxygen culture
conditions enhancing the proliferation, differentiation, and/or viability of mouse, rat, and bovine myoblasts. The specific
effects of low oxygen depend on the myoblast source and oxygen concentration; however, variable oxygen conditions have not
been tested in the culture of human myoblasts. In this study, muscle precursor cells were isolated from vastus lateralis muscle biopsies and myoblast cultures were established in 5% oxygen, before being divided into physiological (5%) or standard
(20%) oxygen conditions for experimental analysis. Five percent oxygen increased proliferating myoblast numbers, and since
low oxygen had no significant effect on myoblast viability, this increase in cell number was attributed to enhanced proliferation.
The proportion of cells in the S (DNA synthesis) phase of the cell cycle was increased by 50%, and p21Cip1 gene and protein expression was decreased in 5 versus 20% oxygen. Unlike in rodent and bovine myoblasts, the increase in
myoD, myogenin, creatine kinase, and myosin heavy chain IIa gene expression during differentiation was similar in 5 and 20%
oxygen; as was myotube hypertrophy. These data indicate for the first time that low oxygen culture conditions stimulate proliferation,
whilst maintaining (but not enhancing) the viability and the differentiation potential of human primary myoblasts and should
be considered as optimum conditions for ex-vivo expansion of these cells. |
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Keywords: | Human primary myoblasts Oxygen Proliferation p21Cip1 Differentiation Cell cycle Species differences |
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