Convenient and Efficient Enrichment of the CD133+Liver Cells from Rat Fetal Liver Cells as a Source of Liver Stem/Progenitor Cells
Convenient and Efficient Enrichment of the CD133+Liver Cells from Rat Fetal Liver Cells as a Source of Liver Stem/Progenitor Cells
复制标题
方便高效地富集大鼠胎儿肝细胞中的 CD133 肝细胞作为肝干/祖细胞的来源
DOI：
10.1007/s12015-010-9119-4
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发表时间：
2011-03-01
影响因子：
4.8
通讯作者：
Dou, Ke-feng
中科院分区：
文献类型：
--
作者：
Liu, Wei-hui;Li, Ren;Dou, Ke-feng
虽然干细胞通常通过FACS或MACS分离，但它们非常昂贵，并且这些干细胞/祖细胞（LSPC）没有特异性标记。本文采用一种简便、有效的方法富集LSPC。采用Percoll不连续梯度离心法（PDGC）富集大鼠胎肝细胞（FLC）。采用差速胰蛋白酶消化和差速贴壁法（DTDA）纯化培养的FLC。流式细胞术分析显示超过一半的纯化的FLC表达LSPC的替代标记物（CD 117、c-Met、Sca-1、CD 90、CD 49 f和CD 133）。换言之，纯化的FLC是异质的。因此，通过Percoll连续梯度离心（PCGC）将它们依次分层成六个级分。CD 133和CD 49 f表达从级分1到6逐渐降低。在组分1和组分2中，约85%的FLC表达CD 133，AFP和CK-19高表达，G-6-P和ALB低表达，表明FLC为LSPC。本研究将为肝干细胞的分离和肝细胞治疗两个重要的生物学方面做出贡献。
Although the stem cells are commonly isolated by FACS or MACS, they are very expensive and these is no specific marker for liver stem/progentior cells (LSPCs). This paper applied a convenient and efficient method to enrich LSPCs. The fetal liver cells (FLCs) were firstly enriched by Percoll discontinuous gradient centrifugation (PDGC) from the rat fetal liver. Then the FLCs in culture were purified to be homogeneous in size by differential trypsinization and differential adherence (DTDA). Flow cytometric analysis revealed more than half of the purified FLCs expressed alternative markers of LSPCs (CD117, c-Met, Sca-1, CD90, CD49f and CD133). In other words, the purified FLCs were heterogeneous. Therefore, they were sequentially layered into six fractions by Percoll continuous gradient centrifugation (PCGC). Both CD133 and CD49f expressed decreasingly from fraction 1 to 6. In fraction 1 and 2, about 85% FLCs expressed CD133, which were revealed to be LSPCs by high expressions of AFP and CK-19, low expressions of G-6-P and ALB. To conclude, the purity of CD133+LSPCs enriched by combination of PDGC, DTDA and PCGC is close to that obtained by MACS. This study will greatly contribute to two important biological aspects: liver stem cells isolation and liver cell therapy.