Cells Isolated from Adult Human Skeletal Muscle Capable of Differentiating into Multiple Mesodermal Phenotypes
Cells Isolated from Adult Human Skeletal Muscle Capable of Differentiating into Multiple Mesodermal Phenotypes
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DOI：
10.1177/000313489906500106
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发表时间：
1999-01
期刊：
影响因子：
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通讯作者：
John T. Williams;Shiela S. Southerland;J. Souza;Andrew F. Calcutt;R. Cartledge
中科院分区：
文献类型：
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作者：
John T. Williams;Shiela S. Southerland;J. Souza;Andrew F. Calcutt;R. Cartledge
伤口愈合是组织对损伤的反应，导致疤痕形成。组织再生将是更理想的反应。此前，我们已经从鸟类、啮齿动物和兔的骨骼肌中分离出一组细胞，能够分化为多种中胚层表型。目前的实验旨在确定在人类骨骼肌中是否存在类似的细胞群体。从75岁女性截肢的腿部骨骼肌和27岁男性的胸肌酶解分离细胞，在含有10%预选马血清的Eagle‘s最低基本培养液中培养融合，然后胰酶消化，过滤，在7.5%-80°C的二甲基亚砜中缓慢冷冻。细胞解冻后，在相同的培养基加地塞米松(一种非特异性分化剂)培养6周。进行免疫组织化学和免疫组织化学染色。观察到的表型包括具有典型星状形态的干细胞(对照组)、骨骼肌管(抗肌球蛋白)、平滑肌(抗α-肌动蛋白)、骨(von Kossa染色)、软骨(Alcec蓝)和脂肪(苏丹黑B)。这些实验证实了人骨骼肌中存在能够分化为多种中胚层表型的间充质干细胞群体。存在操纵间充质干细胞以实现损伤患者的间质组织适当再生的可能性。
Wound healing is the response of tissue to injury that results in scar formation. Tissue regeneration would be a more ideal response. Previously, we have isolated a population of cells from avian, rodent, and rabbit skeletal muscle capable of differentiating into multiple mesodermal phenotypes. The present experiments were designed to determine whether a similar population of cells exist in human skeletal muscle. Separate cell preparations from skeletal muscle on an amputated leg of a 75-year-old female and the pectoralis muscle of a 27-year-old male were enzymatically dissociated and cultured to confluence in Eagle's minimal essential medium with 10 per cent preselected horse serum, then trypsinized, filtered, and slowly frozen in 7.5 per cent dimethylsulfoxide to -80° C. The cells were thawed and plated with the same media plus dexamethasone (a nonspecific differentiation agent) at 10–10–-10–6 M concentrations for up to 6 weeks. Immunological and histochemical staining assays were performed. Phenotypes observed included stem cells with typical stellate morphology (control), skeletal myotubes (anti-myosin), smooth muscle (anti-a-actin), bone (von Kossa stain), cartilage (Alcec blue), and fat (Sudan black B). These experiments establish the existence of a population of mesenchymal stem cells in human skeletal muscle capable of differentiating into multiple mesodermal phenotypes. The possibility exists of manipulating the mesenchymal stem cells to achieve appropriate regeneration of mesenchymal tissues in the injured patient.