Hall Monica N, Hall John K, Cadwallader Adam B, Pawlikowski Bradley T, Doles Jason D, Elston Tiffany L, Olwin Bradley B
Department of Molecular, Cellular and Developmental Biology, University of Colorado, 347 UCB, Boulder, CO, 80309, USA.
Methods Mol Biol. 2017;1556:237-244. doi: 10.1007/978-1-4939-6771-1_12.
Transplanting adult stem cells provides a stringent test for self-renewal and the assessment of comparative engraftment in competitive transplant assays. Transplantation of satellite cells into mammalian skeletal muscle provided the first critical evidence that satellite cells function as adult muscle stem cells. Transplantation of a single satellite cell confirmed and extended this hypothesis, providing proof that the satellite cell is a bona fide adult skeletal muscle stem cell as reported by Sacco et al. (Nature 456(7221):502-506). Satellite cell transplantation has been further leveraged to identify culture conditions that maintain engraftment and to identify self-renewal deficits in satellite cells from aged mice. Conversion of iPSCs (induced pluripotent stem cells) to a satellite cell-like state, followed by transplantation, demonstrated that these cells possess adult muscle stem cell properties as reported by Darabi et al. (Stem Cell Rev Rep 7(4):948-957) and Mizuno et al. (FASEB J 24(7):2245-2253). Thus, transplantation strategies involving either satellite cells derived from adult muscles or derived from iPSCs may eventually be exploited as a therapy for treating patients with diseased or failing skeletal muscle. Here, we describe methods for isolating dispersed adult mouse satellite cells and satellite cells on intact myofibers for transplantation into recipient mice to study muscle stem cell function and behavior following engraftment .
移植成体干细胞为自我更新以及在竞争性移植试验中评估相对植入情况提供了严格的测试。将卫星细胞移植到哺乳动物骨骼肌中,首次提供了关键证据,证明卫星细胞作为成体肌肉干细胞发挥作用。单个卫星细胞的移植证实并扩展了这一假说,证明卫星细胞是一种真正的成体骨骼肌干细胞,正如萨科等人(《自然》456(7221):502 - 506)所报道的那样。卫星细胞移植已被进一步用于确定维持植入的培养条件,并识别老年小鼠卫星细胞中的自我更新缺陷。将诱导多能干细胞(iPSCs)转化为卫星细胞样状态,然后进行移植,结果表明这些细胞具有成体肌肉干细胞特性,正如达拉比等人(《干细胞研究与报告》7(4):948 - 957)以及水野等人(《美国实验生物学会联合会杂志》24(7):2245 - 2253)所报道的那样。因此,涉及源自成体肌肉或iPSCs的卫星细胞的移植策略最终可能被用作治疗患有病变或功能衰竭骨骼肌患者的一种疗法。在此,我们描述了分离分散的成年小鼠卫星细胞和完整肌纤维上的卫星细胞用于移植到受体小鼠体内的方法,以研究植入后肌肉干细胞的功能和行为。