Brennand Kristen, Huangfu Danwei, Melton Doug
Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America.
PLoS Biol. 2007 Jul;5(7):e163. doi: 10.1371/journal.pbio.0050163. Epub 2007 May 29.
In healthy adult mice, the beta cell population is not maintained by stem cells but instead by the replication of differentiated beta cells. It is not known, however, whether all beta cells contribute equally to growth and maintenance, as it may be that some cells replicate while others do not. Understanding precisely which cells are responsible for beta cell replication will inform attempts to expand beta cells in vitro, a potential source for cell replacement therapy to treat diabetes. Two experiments were performed to address this issue. First, the level of fluorescence generated by a pulse of histone 2B-green fluorescent protein (H2BGFP) expression was followed over time to determine how this marker is diluted with cell division; a uniform loss of label across the entire beta cell population was observed. Second, clonal analysis of dividing beta cells was completed; all clones were of comparable size. These results support the conclusion that the beta cell pool is homogeneous with respect to replicative capacity and suggest that all beta cells are candidates for in vitro expansion. Given similar observations in the hepatocyte population, we speculate that for tissues lacking an adult stem cell, they are replenished equally by replication of all differentiated cells.
在健康成年小鼠中,β细胞群体并非由干细胞维持,而是通过已分化β细胞的复制来维持。然而,尚不清楚所有β细胞对生长和维持的贡献是否相同,因为可能有些细胞进行复制而其他细胞不进行复制。精确了解哪些细胞负责β细胞复制,将为体外扩增β细胞的尝试提供信息,而β细胞体外扩增是细胞替代疗法治疗糖尿病的一个潜在来源。为解决这个问题,进行了两项实验。首先,追踪组蛋白2B-绿色荧光蛋白(H2BGFP)表达脉冲产生的荧光水平随时间的变化,以确定该标记物如何随着细胞分裂而稀释;观察到整个β细胞群体中标记物均匀丢失。其次,完成了对分裂β细胞的克隆分析;所有克隆大小相当。这些结果支持了β细胞库在复制能力方面是同质的这一结论,并表明所有β细胞都是体外扩增的候选对象。鉴于在肝细胞群体中也有类似观察结果,我们推测对于缺乏成体干细胞的组织,它们通过所有分化细胞的复制得到同等补充。