Chao S H, Peshwa M V, Sutherland D E, Hu W S
Department of Surgery, School of Medicine, National Taiwan University, ROC.
Cell Transplant. 1992;1(1):51-60. doi: 10.1177/096368979200100109.
In vitro culture of islets of Langerhans decreases their immunogenicity, presumably by eliminating passenger leukocytes and other Ia+ presenting cells within the islets. Islets cultivated in petri dishes either at 37 degrees C or at 25 degrees C gradually disintegrate during culture in a time-dependent manner which is related to the free-floating condition of the islets. Also, a fraction of the islets disperse as single cells and beta-cell aggregates or adhere to the bottom of the culture dishes. Thus, the retrieval rate of transplantable islets is dampened due to their disintegration and spontaneous dispersion in conventional petri dish cultures. Entrapment of freshly harvested islets of Langerhans in a three-dimensional collagen matrix was studied as an alternative method for islet cultivation. The contraction of collagen fibrils during in vitro culture counteracts the dispersion of islets and helps in maintaining their integrity while in culture. It was observed that the entrapped islets maintain satisfactory morphology, viability, and capability of glucose-dependent insulin secretion for over 2 wk. The oxygen consumption rate and glucose metabolism of these islets was not deranged when entrapped in collagen. Also, the retrieval of islets is easier and more efficient than that observed in conventional culture systems. Our results indicate that culture of islets in three-dimensional collagen gels can potentially develop into an ideal system applicable to clinical transplantation of cultured islets or beta-cell aggregates.
胰岛的体外培养可降低其免疫原性，这可能是通过清除胰岛内的过客白细胞和其他Ia + 呈递细胞实现的。在培养皿中于37℃或25℃培养的胰岛在培养过程中会以时间依赖的方式逐渐解体，这与胰岛的自由漂浮状态有关。此外，一部分胰岛会分散为单个细胞和β细胞聚集体，或附着在培养皿底部。因此，由于可移植胰岛在传统培养皿培养中会解体和自发分散，其回收率受到影响。研究了将新鲜收获的胰岛包埋在三维胶原基质中作为胰岛培养的替代方法。体外培养过程中胶原纤维的收缩可抵消胰岛的分散，并有助于在培养过程中保持其完整性。观察到包埋的胰岛在超过2周的时间内保持了令人满意的形态、活力和葡萄糖依赖性胰岛素分泌能力。当包埋在胶原中时，这些胰岛的耗氧率和葡萄糖代谢并未紊乱。此外，与传统培养系统相比，胰岛的回收更容易、更有效。我们的结果表明，在三维胶原凝胶中培养胰岛有可能发展成为一种适用于培养胰岛或β细胞聚集体临床移植的理想系统。
