Bone A J, Swenne I
In Vitro. 1982 Feb;18(2):141-8. doi: 10.1007/BF02796406.
Free islet cell suspensions were prepared from isolated fetal rat islets using a modified enzyme dispersion technique. The islet cells were dispensed into a culture flask containing microcarriers (Cytodex) suspended in culture medium RPMI 1640 by a slowly rotating bar magnet. Microscopical examination of the beads showed that the islet cells attached and then progressively proliferated on the surface of the beads as a monolayer. A highly sustained release of insulin from the beads to the medium was observed during the 7 d culture period. The functional viability of the cultured islet cells was further demonstrated by the ability of batches of the cell-coated beads to synthesize insulin and to increase the insulin release in response to an acute challenge (16.7 mmol/l glucose plus 5 mmol/l theophylline). The results suggest that bead microcarriers may provide a new approach to monolayer islet cell culture providing functional monolayers, which can easily be transferred to different test systems and further manipulated.
采用改良的酶分散技术从分离的胎鼠胰岛制备游离胰岛细胞悬液。将胰岛细胞接种到含有微载体(Cytodex)的培养瓶中,微载体悬浮于RPMI 1640培养基中,通过缓慢旋转的条形磁铁进行搅拌。对微载体进行显微镜检查发现,胰岛细胞附着在微载体表面,随后逐渐以单层形式增殖。在7天的培养期内观察到胰岛素从微载体向培养基的高度持续释放。通过细胞包被微载体批次合成胰岛素以及对急性刺激(16.7 mmol/l葡萄糖加5 mmol/l茶碱)做出反应增加胰岛素释放的能力,进一步证明了培养的胰岛细胞的功能活力。结果表明,微载体珠可能为单层胰岛细胞培养提供一种新方法,形成功能性单层,可轻松转移到不同的测试系统并进一步操作。