期刊
JOURNAL OF ORTHOPAEDIC TRANSLATION
卷 3, 期 1, 页码 26-33出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jot.2014.07.005
关键词
bone marrow; isolation; mesenchymal stem cells; mouse; protocol
类别
资金
- Hong Kong Government Research Grant Council, General Research Fund [CUHK470813]
- China Shenzhen City Science and Technology Bureau under the Shenzhen City Knowledge Innovation Plan, Basic Research Project [JCYJ20130401171935811]
- SMART program, Lui CheWoo Institute of Innovative Medicine, Faculty of Medicine, The Chinese University of Hong Kong
Mesenchymal stem cells (MSCs) from bone marrow are main cell source for tissue repair and engineering, and vehicles of cell-based gene therapy. Unlike other species, mouse bone marrow derived MSCs (BM-MSCs) are difficult to harvest and grow due to the low MSCs yield. We report here a standardised, reliable, and easy-to-perform protocol for isolation and culture of mouse BM-MSCs. There are five main features of this protocol. (1) After flushing bone marrow out of the marrow cavity, we cultured the cells with fat mass without filtering and washing them. Our method is simply keeping the MSCs in their initial niche with minimal disturbance. (2) Our culture medium is not supplemented with any additional growth factor. (3) Our method does not need to separate cells using flow cytometry or immunomagnetic sorting techniques. (4) Our method has been carefully tested in several mouse strains and the results are reproducible. (5) We have optimised this protocol, and list detailed potential problems and trouble-shooting tricks. Using our protocol, the isolated mouse BM-MSCs were strongly positive for CD44 and CD90, negative CD45 and CD31, and exhibited tri-lineage differentiation potentials. Compared with the commonly used protocol, our protocol had higher success rate of establishing the mouse BM-MSCs in culture. Our protocol may be a simple, reliable, and alternative method for culturing MSCs from mouse bone marrow tissues. Copyright (C) 2014, The Authors. Published by Elsevier (Singapore) Pte Ltd. This is an open access article under the CC BY-NC-ND license
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