In vitro neogenesis of human islets reflects the plasticity of differentiated human pancreatic cells
In vitro neogenesis of human islets reflects the plasticity of differentiated human pancreatic cells
复制标题
DOI：
10.1007/s00125-005-1935-8
复制
发表时间：
2005-11-01
期刊：
影响因子：
8.2
通讯作者：
Otonkoski, T
中科院分区：
文献类型：
--
作者：
Gao, R;Ustinov, J;Otonkoski, T
目的/假设：已经报道了培养的成人胰腺组织中胰岛的新生。胰岛祖细胞被认为是导管细胞。由于先前的实验被许多预先存在的胰岛细胞“污染”，我们研究了它们在胰岛细胞新生中的作用。方法：用不同纯度的胰岛细胞单层培养新鲜人胰岛细胞，用溴脱氧尿苷标记。双免疫荧光染色分析移行细胞。对于纯化的导管细胞培养，预先存在的胰岛在磁性细胞分离系统中被消除。结果：我们证实不到1%的内分泌细胞增殖，主要是在培养的前48小时。然而，10倍以上的细胞通过开始共表达导管标记细胞角蛋白19 （CK19）获得过渡表型。这些细胞在培养1天后占所有内分泌细胞的10%以上，在培养5天后占6%。通过磁性细胞分选，我们剔除了表达神经细胞粘附分子（N-CAM）的细胞，之后我们获得了99.7%的纯非内分泌的富含ck19的细胞群。这些细胞群可以在体外扩增。然而，与原始的混合细胞培养相比，它们的内分泌分化能力严重降低。结论/解释：这些结果表明，在这种培养系统中，胰岛新生至少部分代表了胰岛细胞向导管样表型的去分化，并在适当的条件下进一步再分化。人类胰腺细胞类型分化的可塑性可能是人类胰腺再生的重要机制。
Aims/hypothesis: The neogenesis of islets from cultured human adult pancreatic tissue has been reported. The islet progenitors have been thought to be ductal cells. Since previous experiments have been 'contaminated' by a number of pre-existing islet cells, we examined their involvement in islet cell neogenesis. Methods: Fresh human pancreatic cells with different purities of islet cells were grown in monolayer culture and labelled with bromodeoxyuridine. Transitional cells were analysed by double immunofluorescence staining. For purified ductal cell culture, pre-existing islets were eliminated on a magnetic cell separation system. Results: We confirmed that less than 1% of the endocrine cells proliferated, mainly during the first 48 h of culture. However, a 10-fold larger proportion of the cells acquired a transitional phenotype by starting to coexpress the ductal marker cytokeratin 19 (CK19). These cells represented more than 10% of all endocrine cells after 1 day in culture, and 6% at 5 days of culture. Using magnetic cell sorting, we eliminated cells expressing neural cell adhesion molecule (N-CAM), after which we obtained 99.7% pure non-endocrine CK19-rich cell populations. These cell populations could be expanded in vitro. However, their endocrine differentiation capacity was severely reduced as compared with the original mixed cell cultures. Conclusions/interpretation: These results suggest that islet neogenesis in this culture system at least partly represents the de-differentiation of islet cells into a duct-cell-like phenotype, with further re-differentiation in appropriate conditions. The plasticity of differentiated human pancreatic cell types may thus be an important mechanism of human pancreas regeneration.