Okumachi Etsuko, Lee Sang Yang, Niikura Takahiro, Iwakura Takashi, Dogaki Yoshihiro, Waki Takahiro, Takahara Shunsuke, Ueha Takeshi, Sakai Yoshitada, Kuroda Ryosuke, Kurosaka Masahiro
Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, 7-5-1, Kusunoki-cho, Chuo-ku, Kobe, Japan, 650-0017.
Int Orthop. 2015 Mar;39(3):569-76. doi: 10.1007/s00264-014-2569-6. Epub 2014 Oct 23.
Skeletal muscle comprises different kinds of muscle fibres that can be classified as slow and fast fibres. The purpose of this study was to compare the yield, proliferation, and multi-potentiality of rat mesenchymal stem cells (MSCs) from the tibialis anterior (TA; fast muscle) and soleus (SO; slow muscle) in vitro.
The TA and SO muscles were harvested, and isolated cells were plated. After two hours, the cells were washed extensively to remove any cell that did not adhere to the cell culture plate. The adherent cells, namely MSCs, were then cultured. Both types of MSCs were differentiated toward the osteogenic, chondrogenic and adipogenic lineages using lineage specific induction factors.
The colony-forming unit fibroblast (CFU-F) assay revealed that the SO contained significantly higher quantities of MSCs than the TA. The self-renewal capacity of MSCs derived from the TA was significantly higher at later passages (passage 9-11). Both types of MSCs exhibited similar cell surface antigens to bone marrow (BM)-derived MSCs and were positive for CD29, CD44, and CD90 and negative for CD11b, CD34, and CD45. TA-derived MSCs were superior in terms of osteogenic differentiation capacity, but there was no significant difference in chondrogenic and adipogenic differentiation capacity.
Our results demonstrated significant differences in the properties of muscle-derived MSCs from different muscle types (i.e. fast or slow muscles). The greater expandability and osteogenic differentiation ability of TA-derived MSCs suggests that fast muscle may be a better source for generating large numbers of MSCs for bone regeneration.
骨骼肌由不同类型的肌纤维组成,可分为慢肌纤维和快肌纤维。本研究的目的是比较大鼠胫骨前肌(TA;快肌)和比目鱼肌(SO;慢肌)来源的间充质干细胞(MSC)在体外的产量、增殖能力和多能性。
采集TA和SO肌肉,将分离出的细胞接种。两小时后,对细胞进行充分洗涤以去除未粘附在细胞培养板上的任何细胞。然后培养贴壁细胞,即MSC。使用谱系特异性诱导因子将两种类型的MSC分别诱导分化为成骨、成软骨和脂肪生成谱系。
集落形成单位成纤维细胞(CFU-F)分析显示,SO中MSC的含量明显高于TA。在传代后期(第9 - 11代),TA来源的MSC的自我更新能力明显更高。两种类型的MSC均表现出与骨髓(BM)来源的MSC相似的细胞表面抗原,CD29、CD44和CD90呈阳性,CD11b、CD34和CD45呈阴性。TA来源的MSC在成骨分化能力方面更优,但在成软骨和成脂分化能力方面无显著差异。
我们的结果表明,不同肌肉类型(即快肌或慢肌)来源的肌肉衍生MSC的特性存在显著差异。TA来源的MSC具有更强的扩增能力和成骨分化能力,这表明快肌可能是用于骨再生的大量MSC的更好来源。