Cells with surface expression of CD133highCD71low are enriched for tripotent colony-forming progenitor cells in the adult murine pancreas.
Cells with surface expression of CD133highCD71low are enriched for tripotent colony-forming progenitor cells in the adult murine pancreas.
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DOI：
10.1016/j.scr.2015.11.015
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发表时间：
2016-01
影响因子：
1.2
通讯作者：
Ku HT
中科院分区：
文献类型：
--
作者：
Jin L;Gao D;Feng T;Tremblay JR;Ghazalli N;Luo A;Rawson J;Quijano JC;Chai J;Wedeken L;Hsu J;LeBon J;Walker S;Shih HP;Mahdavi A;Tirrell DA;Riggs AD;Ku HT
成人胰腺中的祖细胞是治疗 1 型糖尿病的内分泌 β 细胞的潜在来源。此前，我们在成年（2-4月龄）小鼠胰腺中鉴定出三能祖细胞，它们能够在体外自我更新并分化为导管细胞、腺泡细胞和内分泌细胞。这些祖细胞被命名为胰腺集落形成单位（PCFU）。然而，由于 PCFU 是胰腺中的一小部分 (~1%)，因此很难研究。为了丰富 PCFU，开发了使用细胞表面标记分析和荧光激活细胞分选的策略。我们发现，与未分选的细胞相比，CD133highCD71low 细胞（而非其他细胞群）的 PCFU 富集程度高达 30 倍。当连续重新接种到含有基质胶的培养物中时，CD133highCD71low 细胞产生初级、次级和后续集落，表明其具有自我更新能力。在存在层粘连蛋白水凝胶的情况下，CD133highCD71low细胞产生包含导管、腺泡和胰岛素+胰高血糖素+双激素内分泌细胞的集落。将层粘连蛋白水凝胶培养物的集落植入糖尿病小鼠体内，五周后在移植物中检测到导管、腺泡和胰岛素+胰高血糖素细胞，证明了 CD133highCD71low 细胞的三系分化潜力。这些 CD133highCD71low 细胞将使未来对假定的成体胰腺干细胞的体内研究成为可能。
Progenitor cells in the adult pancreas are potential sources of endocrine beta cells for treating type 1 diabetes. Previously, we identified tri-potent progenitor cells in the adult (2–4 month-old) murine pancreas that were capable of self-renewal and differentiation into duct, acinar, and endocrine cells in vitro. These progenitor cells were named pancreatic colony-forming units (PCFUs). However, because PCFUs are a minor population in the pancreas (~1%) they are difficult to study. To enrich PCFUs, strategies using cell-surface marker analyses and fluorescence-activated cell sorting were developed. We found that CD133highCD71low cells, but not other cell populations, enriched PCFUs by up to 30 fold compared to the unsorted cells. CD133highCD71low cells generated primary, secondary, and subsequent colonies when serially re-plated in Matrigel-containing cultures, suggesting self-renewal abilities. In the presence of a laminin hydrogel, CD133highCD71low cells gave rise to colonies that contained duct, acinar, and Insulin+Glucagon+ double-hormonal endocrine cells. Colonies from the laminin hydrogel culture were implanted into diabetic mice, and five weeks later duct, acinar, and Insulin+Glucagon− cells were detected in the grafts, demonstrating tri-lineage differentiation potential of CD133highCD71low cells. These CD133highCD71low cells will enable future studies of putative adult pancreas stem cells in vivo.