Ju M K, Jeong J H, Lee J I, Kim Y S, Kim M S
Research Institute for Transplantation, Yonsei University Health System, Seoul, South Korea; Department of Transplantation Surgery, Yonsei University Health System, Seoul, South Korea.
Transplant Proc. 2013 Jun;45(5):1885-8. doi: 10.1016/j.transproceed.2012.12.029.
Many obstacles beset islet transplantation, particularly insufficient tissue mass. Previously, we reported production of pseudo-islets. In addition, there have been reports in which coculture with pancreatic islet and bone marrow mesenchymal stem cells (BMSCs) demonstrated positive effects on pancreatic islet function. The purpose of this study was to perform morphologic and functional evaluations of pancreatic pseudo-islets cocultured with BMSCs. Pancreatic endocrine cells (PECs) were collected with a previously reported method; bone marrow was aspirated from the rat femur. Subsequently, PECs and BMSCs cocultured at high density on low-cell-binding culture dishes kept suspended by shaking. The functionality and characteristics of the mixed cell complexes were evaluated by glucose challenge, insulin enzyme-linked immunosorbent assay, reverse-transcription polymerase chain reaction, and immunohistochemistry. Through expansion for 2 weeks in continuous culture passages, ∼1 million PECs were recovered after aggregation. They presented spherical shapes and sizes similar to naïve islets, according to phase-contrast microscopy. The spheroid aggregates of pancreatic islet cells and BMSCs showed fortified functions and maintained viability. In conclusion, PECs served as a cell source for pseudo-islets, which were both morphologically and genetically similar to naïve islets. We also suggest a manufacturing method for mixed cellular complexes from 2 different origins that can improve secretion ability and cell differentiation.
胰岛移植面临诸多障碍,尤其是组织量不足。此前,我们报道了伪胰岛的产生。此外,有报道称胰腺胰岛与骨髓间充质干细胞(BMSCs)共培养对胰岛功能有积极影响。本研究的目的是对与BMSCs共培养的胰腺伪胰岛进行形态学和功能评估。采用先前报道的方法收集胰腺内分泌细胞(PECs);从大鼠股骨中抽取骨髓。随后,将PECs和BMSCs在低细胞结合培养皿上高密度共培养,并通过振荡使其保持悬浮状态。通过葡萄糖刺激、胰岛素酶联免疫吸附测定、逆转录聚合酶链反应和免疫组织化学对混合细胞复合物的功能和特性进行评估。经过连续传代培养2周的扩增,聚集后回收了约100万个PECs。根据相差显微镜观察,它们呈现出与天然胰岛相似的球形和大小。胰腺胰岛细胞和BMSCs的球体聚集体显示出强化的功能并保持了活力。总之,PECs作为伪胰岛的细胞来源,在形态和基因上均与天然胰岛相似。我们还提出了一种由两种不同来源制造混合细胞复合物的方法,该方法可提高分泌能力和细胞分化。