Hematopoietic potential of murine skeletal muscle-derived CD45-Sca-1+c-kit- cells
Hematopoietic potential of murine skeletal muscle-derived CD45-Sca-1+c-kit- cells
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DOI：
10.1016/s0301-472x(02)00872-x
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发表时间：
2002-08-01
影响因子：
2.6
通讯作者：
Srour, EF
中科院分区：
文献类型：
--
作者：
Howell, JC;Yoder, MC;Srour, EF
目标。体细胞在出生后的哺乳动物组织中没有明确的定义，是研究干细胞可塑性的有吸引力的候选细胞。我们的目标是确定具有造血潜力的新生儿肌肉来源细胞的身份，并探索这些细胞上CD45的表达状况。材料和方法。取4-7日龄小鼠大腿骨骼肌，酶消化，流式细胞仪分选，得到CD45(-)、Sca-1(+)、c-kit(-)细胞。这些细胞在造血祖细胞集落形成试验和竞争性再繁殖试验中进行检测，并进行体外扩增。此外，随着时间的推移，追踪CD45、c-kit、PU.1和β珠蛋白在培养细胞中的主要表达，以评估在体外操纵干细胞分化的可能性。新鲜分离的CD45(-)SCA-1(+)c-kit(-)细胞缺乏造血系标志，不含集落形成活性，但与新鲜分离的肌肉来源的CD45(+)SCA-1(+)c-kit(+)细胞相比，显示出更好的长期竞争性再扩增能力。CD45(-)SCA-1(+)c-kit(-)细胞在5 ng/mL小鼠干细胞因子mFlt-3L和巨核细胞生长发育因子(MGDF)体外扩增9天后，其体内造血再扩增能力是新鲜细胞的5.3倍。尽管新鲜细胞不转录多种造血基因的mRNA，但培养9d的细胞中有一小部分获得了细胞表面的c-kit，只有这些细胞表达了c-kit和PU.1基因，并保持了竞争性的再扩增能力，表明至少具有髓系和淋巴系的发育潜力。表达CD45(-)、Sca-1(+)、c-kit(-)表型的新生小鼠肌源性细胞是体外和体内具有造血分化潜能的成体干细胞。(C)2002年国际实验血液学学会。爱思唯尔科学公司出版。
Objective. Somatic stem cells, which are poorly defined in postnatal mammalian tissues, are attractive candidates for examination of stem cell plasticity. Our goal was to determine the identity of neonatal muscle-derived cells that contain hematopoietic potential and to explore the status of CD45 expression on these cells.Materials and Methods. Skeletal muscle from thighs of 4- to 7-day-old mice was harvested, enzymatically digested, and flow cytometrically sorted to yield CD45(-)Sca-1(+)c-kit(-) cells. These cells were examined in hematopoietic colony-forming assays and competitive repopulation assays, and were expanded ex vivo. Additionally, CD45, c-kit, PU.1, and beta globin major expression was tracked over time in cultured cells to assess the possibility of manipulating stem cell differentiation in vitro.Results. Freshly isolated CD45(-)Sca-1(+)c-kit(-) cells were devoid of hematopoietic lineage markers and contained no colony-forming activity but displayed superior long-term competitive repopulating ability when compared to freshly isolated muscle-derived CD45(+)Sca-1(+)c-kit(+) cells. CD45(-)Sca-1(+)c-kit(-) cells expanded ex vivo in 5 ng/mL murine stem cell factor, mFlt-3L, and megakaryocyte growth and development factor (MGDF) for 9 days increased their in vivo hematopoietic repopulating potential 5.3-fold relative to fresh cells. Although fresh cells did not transcribe mRNA of several hematopoietic genes, a small fraction of cells cultured for 9 days acquired cell surface c-kit, and only these cells expressed c-kit and PU.1 mRNA and maintained competitive repopulating ability, suggesting at least myeloid and perhaps lymphoid developmental potential.Conclusion. Neonatal murine muscle-derived cells expressing the phenotype CD45(-)Sca-1(+) c-kit(-) are putative adult somatic stem cells with in vitro and in vivo hematopoietic differentiation potential. (C) 2002 International Society for Experimental Hematology. Published by Elsevier Science Inc.