Background

The use of pancreata from pre-weaned piglets has the potential to serve as an unlimited alternative source of islets for clinical xenotransplantation. As pre-weaned porcine islets (PPIs) are immature and require prolonged culture, we developed an islet maturation media (IMM) and evaluated its effect on improving the quantity and quality of PPIs over 14 days of culture.

Methods

PPIs were isolated from the pancreata of pre-weaned Yorkshire piglets (8–15 days old). Each independent islet isolation was divided for culture in either control Ham’s F-10 media (n = 5) or IMM (n = 5) for 14 days. On day 3, 7 and 14 of culture, islets were assessed for islet yield, isolation index, viability, insulin content, endocrine cellular composition, differentiation of beta cells, and insulin secretion during glucose stimulation.

Results

In comparison to control islets, culturing PPIs in IMM significantly increased islet yield. PPIs cultured in IMM also maintained a stable isolation index and viability throughout 14 days of culture. The insulin content, endocrine cellular composition, and differentiation of beta cells were significantly improved in PPIs cultured in IMM, which subsequently augmented their insulin secretory capacity in response to glucose challenge compared to control islets.

Conclusions

Culturing PPIs in IMM increases islet yield, isolation index, viability, insulin content, endocrine cellular composition, differentiation of endocrine progenitor cells toward beta cells, and insulin secretion. Due to the improved islet quantity and quality after in vitro culture, the use of IMM in the culture of PPIs will assist to advance the outcomes of clinical islet xenotransplantation.

中文翻译：

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一种胰岛成熟培养基，可在体外培养过程中改善年轻猪胰岛的发育。

背景

使用断奶前仔猪的胰腺有可能作为临床异种移植的无限替代胰岛来源。由于断奶前猪胰岛 (PPI) 不成熟且需要长时间培养，我们开发了胰岛成熟培养基 (IMM) 并评估了其在培养 14 天后对提高 PPI 数量和质量的影响。

方法

从断奶前约克夏仔猪（8-15 日龄）的胰腺中分离出 PPI。每个独立的胰岛分离物在对照火腿 F-10 培养基 (n = 5) 或 IMM (n = 5) 中分开培养 14 天。在培养的第 3、7 和 14 天，评估胰岛的胰岛产量、分离指数、活力、胰岛素含量、内分泌细胞组成、β 细胞分化和葡萄糖刺激期间的胰岛素分泌。

结果

与对照胰岛相比，在 IMM 中培养 PPI 显着增加了胰岛产量。在 IMM 中培养的 PPI 在整个 14 天的培养过程中也保持了稳定的分离指数和活力。在 IMM 中培养的 PPI 中，胰岛素含量、内分泌细胞组成和 β 细胞的分化得到显着改善，与对照胰岛相比，随后增加了它们响应葡萄糖挑战的胰岛素分泌能力。

结论

在 IMM 中培养 PPI 可增加胰岛产量、分离指数、活力、胰岛素含量、内分泌细胞组成、内分泌祖细胞向 β 细胞的分化和胰岛素分泌。由于体外培养后胰岛数量和质量的提高，在 PPI 培养中使用 IMM 将有助于推进临床胰岛异种移植的结果。