Protection from cell death in cultured human fetal pancreatic cells
Protection from cell death in cultured human fetal pancreatic cells
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DOI：
10.1177/096368970000900314
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发表时间：
2000-05-01
影响因子：
3.3
通讯作者：
Hayek, A
中科院分区：
文献类型：
--
作者：
Beattie, GM;Leibowitz, G;Hayek, A
来自人胎儿胰腺的内分泌细胞将在体外在细胞外基质上增殖，但失去激素表达，并且再分化需要从基质中去除扩增的细胞并重新聚集成细胞聚集体。然而，在操作和培养过程中会发生大量细胞死亡。在不同的实验条件下，在整个过程的每个阶段检查细胞死亡的机制，以确定提高细胞活力的最佳方案。在运输期间，向培养基中添加海藻糖以防止坏死使产量增加17倍，而在培养胰岛样细胞簇期间，凋亡抑制剂Z-VAD使产量增加1.8倍。在细胞-基质相互作用和再聚集的破坏之后，通过TUNEL测定有明显的凋亡小体的证据。添加烟酰胺或Z-VAD，或在再聚集过程中从培养基中去除精氨酸，减少了凋亡小体的数量，并且效果是累加的。然而，需要组合处理以显著增加活细胞的产量。我们的结论是，细胞死亡的人胎儿胰腺组织在文化中的结果从坏死和凋亡，并在细胞水平上的机制的理解将导致协议，将提高细胞活力，促进β-细胞生长。
Endocrine cells from the human fetal pancreas will proliferate in vitro on extracellular matrix but lose hormone expression, and redifferentiation requires removal of the expanded cells from the matrix and reaggregation into cell aggregates. However, extensive cell death occurs during manipulation and culture. The mechanism of cell death was examined at each stage throughout the process under different experimental conditions to determine optimal protocols to increase cell viability. During shipment, the addition of trehalose to the media to prevent necrosis increased yield 17-fold, while during culture as islet-like cell clusters the apoptosis inhibitor Z-VAD increased yield 1.8-fold. Following disruption of cell-matrix interactions and reaggregation, there was marked evidence of apoptotic bodies by the TUNEL assay. Addition of nicotinamide or Z-VAD, or removal of arginine from the media during reaggregation, reduced the number of apoptotic bodies and the effect was additive. However, a combination of treatments was necessary to significantly increase the yield of viable cells. We conclude that cell death of human fetal pancreatic tissue in culture results from both necrosis and apoptosis and that understanding the mechanisms at the cellular level will lead to protocols that will improve cell viability and promote beta-cell growth.