Baylor All Saints Medical Center, Baylor Research Institute, Fort Worth, TX 76104, USA.
Cell Transplant. 2010;19(6):879-86. doi: 10.3727/096368910X509004. Epub 2010 Jun 29.
β-Cell replacement therapy via islet transplantation is an effective treatment for diabetes mellitus, but its widespread use is severely limited by the shortage of donor organs. Because pancreatic stem/progenitor cells are abundantly available in the pancreas of these patients and in donor organs, the cells could become a useful target for β-cell replacement therapy. We previously established a mouse pancreatic stem cell line without genetic manipulation. In this study, we used the techniques to identify and isolate human pancreatic stem/progenitor cells. The cells from a duct-rich population were cultured in 23 kinds of culture media, based on media for mouse pancreatic stem cells or for human embryonic stem cells. The cells in serum-free media formed "cobblestone" morphologies, similar to a mouse pancreatic stem cell line. On the other hand, the cells in serum-containing medium and the medium for human embryonic stem cells formed "fibroblast-like" morphologies. The cells divided actively until day 30, and the population doubling level (PDL) was 6-10. However, the cells stopped dividing after 30 days in any culture conditions. During the cultures, the nucleus/cytoplasm (N/C) ratio decreased, suggesting that the cells entered senescence. Exendin-4 treatment and transduction of PDX-1 and NeuroD proteins by protein transduction technology into the cells induced insulin and pancreas-related gene expression. Although the duplications of these cells were limited, this approach could provide a potential new source of insulin-producing cells for transplantation.
β细胞替代治疗通过胰岛移植是一种有效的治疗糖尿病的方法，但由于供体器官的短缺，其广泛应用受到严重限制。由于这些患者和供体器官的胰腺中富含胰腺干细胞/祖细胞，因此这些细胞可能成为β细胞替代治疗的有用目标。我们之前建立了一种无需遗传操作的小鼠胰腺干细胞系。在这项研究中，我们使用了鉴定和分离人胰腺干细胞/祖细胞的技术。从富含导管的细胞群中，我们在 23 种培养基中培养这些细胞，这些培养基基于小鼠胰腺干细胞或人胚胎干细胞的培养基。无血清培养基中的细胞形成“鹅卵石”形态，类似于小鼠胰腺干细胞系。另一方面，含血清培养基和人胚胎干细胞的培养基中的细胞形成“成纤维细胞样”形态。细胞在 30 天内积极分裂，群体倍增水平（PDL）为 6-10。然而，在任何培养条件下，细胞在 30 天后停止分裂。在培养过程中，核/细胞质（N/C）比值降低，表明细胞进入衰老。Exendin-4 处理和蛋白转导技术将 PDX-1 和 NeuroD 蛋白转导到细胞中诱导胰岛素和胰腺相关基因表达。尽管这些细胞的复制受到限制，但这种方法可以为移植提供潜在的新的胰岛素产生细胞来源。
