Self-Renewing Tri-Potent Stem/Progenitor-like Cells from Adult Human Pancreas

Stem and progenitor cells in the adult human pancreas provide an under-explored resource for regenerative medicine. Using micro-manipulation and methylcellulose-containing colony/organoid assays, we identified cells within the human cadaveric exocrine pancreas that fulfill the definition of a stem cell: able to self-renew and differentiate. Exocrine tissues were collected after the isolation of endocrine cells, dissociated into single cells, and plated into a 3-dimensional semisolid medium. We found that some pancreatic ductal cells gave rise to cystic colonies/organoids containing pancreatic duct, acinar, and endocrine lineage cells. These cells self-renewed and expanded approximately 300-fold over 9 weeks. When transplanted into diabetic mice, colonies/organoids lowered blood glucose levels and gave rise to insulin-expressing endocrine cells. Thus, stem/progenitor-like cells capable of self-renewal and differentiation either preexist in the adult human pancreas or readily adapt in culture. These human cells have implications for therapy in diabetes patients. Total RNAs were extracted from individual cells obtained following dissociation of freshly obtained human adult islet-depleted pancreas tissue.

成人胰腺中的干/祖细胞（stem and progenitor cells）是再生医学领域尚未被充分探索的宝贵研究资源。本研究借助显微操作（micro-manipulation）技术与含甲基纤维素（methylcellulose）的集落/类器官（colony/organoid）培养体系，从人类尸体来源的外分泌胰腺（exocrine pancreas）中筛选出符合干细胞（stem cell）定义的细胞：此类细胞具备自我更新与多向分化能力。研究人员在分离内分泌细胞（endocrine cells）后收集外分泌组织，将其解离为单个细胞并接种于三维半固体培养基（3-dimensional semisolid medium）中。实验结果显示，部分胰腺导管细胞可形成囊性集落/类器官，其中包含胰腺导管、腺泡（acinar）与内分泌谱系（endocrine lineage）细胞。这类细胞可实现自我更新，并在9周内实现约300倍的扩增。将上述集落/类器官移植入糖尿病小鼠体内后，可有效降低小鼠血糖水平，并分化为表达胰岛素（insulin）的内分泌细胞。综上，成人胰腺中要么预先存在具备自我更新与分化能力的干/祖细胞样细胞，要么可在体外培养环境中快速适应并获得该特性。此类人类细胞可为糖尿病患者的治疗提供潜在的应用思路。本研究从新鲜获取的、去除胰岛的成人胰腺组织解离得到的单个细胞中提取了总RNA（total RNAs）。