Andrew A, Rawdon B B, Alison B C
Department of Anatomy and Human Biology, Medical School, University of the Witwatersrand, Johannesburg, South Africa.
In Vitro Cell Dev Biol Anim. 1994 Oct;30A(10):664-70. doi: 10.1007/BF02631269.
Little being known about factors necessary for insulin cell differentiation, we tested the chance observation that these cells were virtually absent from collagen gel cultures of embryonic avian pancreas in which the other pancreatic endocrine cells were numerous. Five-day dorsal buds stripped of their enveloping mesenchyme were embedded in gel and overlaid by a defined medium containing serum, then cultured for 7 days. Immunocytochemical evaluation showed a very low proportion of insulin cells. Substitution of the gel by a polyamino acid coating slightly increased the proportion. In an attempt to test for ability of insulin cell formation to recover, we transferred explants first cultured in collagen gel to polyamino-acid-coated dishes for a further 7 days. No improvement resulted. In controls grown for 14 days on a polyamino acid coating, insulin cells disappeared completely. We conclude that collagen gel does not support survival and differentiation of chick embryonic insulin cells and that the medium used is lacking in some essential factor(s). Determination of their identity should prove possible by exploitation of this model.
由于对胰岛素细胞分化所需的因素了解甚少,我们对一个偶然的观察结果进行了验证:在胚胎期鸡胰腺的胶原凝胶培养物中,这些细胞几乎不存在,而其他胰腺内分泌细胞却大量存在。去除包膜间充质的5日龄背芽被嵌入凝胶中,并用含有血清的特定培养基覆盖,然后培养7天。免疫细胞化学评估显示胰岛素细胞的比例非常低。用聚氨基酸涂层替代凝胶可使比例略有增加。为了测试胰岛素细胞形成恢复的能力,我们将最初在胶原凝胶中培养的外植体转移到聚氨基酸涂层培养皿中再培养7天。但没有得到改善。在聚氨基酸涂层上生长14天的对照中,胰岛素细胞完全消失。我们得出结论,胶原凝胶不支持鸡胚胎胰岛素细胞的存活和分化,并且所用的培养基缺乏某些必需因子。利用这个模型应该有可能确定它们的身份。