High-Yield Purification, Preservation, and Serial Transplantation of Human Satellite Cells.
High-Yield Purification, Preservation, and Serial Transplantation of Human Satellite Cells.
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DOI：
10.1016/j.stemcr.2018.01.022
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发表时间：
2018-03-13
影响因子：
5.9
通讯作者：
Pomerantz JH
中科院分区：
文献类型：
--
作者：
Garcia SM;Tamaki S;Lee S;Wong A;Jose A;Dreux J;Kouklis G;Sbitany H;Seth R;Knott PD;Heaton C;Ryan WR;Kim EA;Hansen SL;Hoffman WY;Pomerantz JH
人类肌肉再生的研究需要强有力的方法来纯化和移植共同构成人类卫星细胞（HuSC）库的肌肉干细胞和祖细胞。现有方法尚未使研究人员能够广泛使用 HuSC，因此人类肌肉干细胞研究进展缓慢。在这里，我们描述了一种稳健且可预测的 HuSC 纯化过程，该过程对测试的每个人类骨骼肌都有效，并在肌营养不良蛋白缺陷和野生型受体中开发了储存方案和移植模型。酶消化、磁力柱去除和 6 标记流式细胞术纯化能够分离每克肌肉 104 个高度富集的 HuSC。 HuSC 的冷冻保存可保留活力、表型和移植潜力。增强型和物种特异性移植方案的开发使得连续 HuSC 异种移植和恢复成为可能。这些协议和模型为 HuSC 的基础和转化研究以及临床开发提供了一个可访问的系统。高效纯化允许连续移植人类卫星干细胞冷冻保存保留卫星细胞功能和表型1克成人骨骼肌产生104个高度纯化的卫星细胞纯化的未培养的内源性人类卫星细胞可以存储和共享Garcia及其同事报告了从人类骨骼肌中有效纯化卫星细胞的方法。他们利用他们的方法来展示内源卫星细胞的干细胞功能，并使人类卫星细胞可供研究人员共享。
Investigation of human muscle regeneration requires robust methods to purify and transplant muscle stem and progenitor cells that collectively constitute the human satellite cell (HuSC) pool. Existing approaches have yet to make HuSCs widely accessible for researchers, and as a result human muscle stem cell research has advanced slowly. Here, we describe a robust and predictable HuSC purification process that is effective for each human skeletal muscle tested and the development of storage protocols and transplantation models in dystrophin-deficient and wild-type recipients. Enzymatic digestion, magnetic column depletion, and 6-marker flow-cytometric purification enable separation of 104 highly enriched HuSCs per gram of muscle. Cryostorage of HuSCs preserves viability, phenotype, and transplantation potential. Development of enhanced and species-specific transplantation protocols enabled serial HuSC xenotransplantation and recovery. These protocols and models provide an accessible system for basic and translational investigation and clinical development of HuSCs. High-efficiency purification permits serial transplantation of human satellite stem cells Cryopreservation preserves satellite cell function and phenotype 1 gram of adult skeletal muscle yields 104 highly purified satellite cells Purified uncultured endogenous human satellite cells can be stored and shared Garcia and colleagues report methods for efficient purification of satellite cells from human skeletal muscle. They use their approaches to demonstrate stem cell functions of endogenous satellite cells and to make human satellite cells accessible for sharing among researchers.
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影响因子：
64.8
作者：
Sacco, Alessandra;Doyonnas, Regis;Kraft, Peggy;Vitorovic, Stefan;Blau, Helen M.
通讯作者：
Blau, Helen M.
影响因子：
64.5
作者：
Sherwood, RI;Christensen, JL;Wagers, AJ
通讯作者：
Wagers, AJ
影响因子：
4.1
作者：
Chirieleison, Steven M.;Feduska, Joseph M.;Deasy, Bridget M.
通讯作者：
Deasy, Bridget M.
体内扩张和体内自我更新人类肌肉干细胞。
影响因子：
5.9
作者：
Charville GW;Cheung TH;Yoo B;Santos PJ;Lee GK;Shrager JB;Rando TA
通讯作者：
Rando TA
影响因子：
82.9
作者：
Dumont NA;Wang YX;von Maltzahn J;Pasut A;Bentzinger CF;Brun CE;Rudnicki MA
通讯作者：
Rudnicki MA