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多能性猪骨骼肌卫星细胞的分离、培养及生物学特性

Isolation, culture and biological characteristics of multipotent porcine skeletal muscle satellite cells.

作者信息

Yang Jinjuan, Liu Hao, Wang Kunfu, Li Lu, Yuan Hongyi, Liu Xueting, Liu Yingjie, Guan Weijun

机构信息

Department of Animal Genetic Resources (AnGR), Institute of Animal Science, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan West Road, Haidian District, Beijing, 100193, People's Republic of China.

Sports Science College, Beijing Sports University, Beijing, 100084, People's Republic of China.

出版信息

Cell Tissue Bank. 2017 Dec;18(4):513-525. doi: 10.1007/s10561-017-9614-9. Epub 2017 Mar 2.

Abstract

Skeletal muscle has a huge regenerative potential for postnatal muscle growth and repair, which mainly depends on a kind of muscle progenitor cell population, called satellite cell. Nowadays, the majority of satellite cells were obtained from human, mouse, rat and other animals but rarely from pig. In this article, the porcine skeletal muscle satellite cells were isolated and cultured in vitro. The expression of surface markers of satellite cells was detected by immunofluorescence and RT-PCR assays. The differentiation capacity was assessed by inducing satellite cells into adipocytes, myoblasts and osteoblasts. The results showed that satellite cells isolated from porcine tibialis anterior were subcultured up to 12 passages and were positive for Pax7, Myod, c-Met, desmin, PCNA and NANOG but were negative for Myogenin. Satellite cells were also induced to differentiate into adipocytes, osteoblasts and myoblasts, respectively. These findings indicated that porcine satellite cells possess similar biological characteristics of stem cells, which may provide theoretical basis and experimental evidence for potential therapeutic application in the treatment of dystrophic muscle and other muscle injuries.

摘要

骨骼肌对于出生后肌肉的生长和修复具有巨大的再生潜力,这主要依赖于一种称为卫星细胞的肌肉祖细胞群体。如今,大多数卫星细胞是从人、小鼠、大鼠和其他动物中获取的,而从猪中获取的却很少。在本文中,猪骨骼肌卫星细胞被分离并进行体外培养。通过免疫荧光和RT-PCR检测卫星细胞表面标志物的表达。通过将卫星细胞诱导分化为脂肪细胞、成肌细胞和成骨细胞来评估其分化能力。结果表明,从猪胫骨前肌分离的卫星细胞可传代培养至12代,且Pax7、Myod、c-Met、结蛋白、增殖细胞核抗原(PCNA)和NANOG呈阳性,但生肌调节因子呈阴性。卫星细胞也分别被诱导分化为脂肪细胞、成骨细胞和成肌细胞。这些发现表明,猪卫星细胞具有与干细胞相似的生物学特性,这可能为治疗营养不良性肌肉和其他肌肉损伤的潜在治疗应用提供理论依据和实验证据。

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