Insulin improves survival but does not maintain function of cultured chick wing bud apical ectodermal ridge.
Insulin improves survival but does not maintain function of cultured chick wing bud apical ectodermal ridge.
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胰岛素可以提高存活率，但不能维持培养鸡翅芽顶端外胚层脊的功能。
DOI：
10.1002/ar.1092330314
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Fallon,JF
中科院分区：
文献类型：
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作者：
Boutin,EL;Fallon,JF
之前我们证明，高水平的胰岛素（5 μg/ml）可以使离体的鸡顶外胚层嵴在培养物中存活（Boutin and Fallon, Dev. Biol., 104:111–116, 1984）。在这里，我们探讨较低水平的胰岛素或胰岛素样生长因子（IGF）是否也可以提高培养的顶端外胚层嵴的存活率，以及嵴功能是否随着嵴存活而得以维持。 IGF I 和 IGF II (100 ng/ml) 均不会减少脊细胞死亡；然而，50 ng/ml 胰岛素可显着减少细胞死亡。在 IGF I 和胰岛素同时存在的情况下，没有获得进一步的改善。这些数据表明胰岛素通过结合其自身受体提高了分离的顶端外胚层嵴的存活率。为了测试功能的维持情况，在有或没有胰岛素（5 μg/ml 或 5 ng/ml）的情况下将第 20 阶段的脊培养 0、6、12、18 或 24 小时，并用于制造重组肢体。培养 12 小时或更长时间的孤立脊产生较少的生长物，并且这些生长物很少在远端完整。尽管细胞存活率存在重大差异，但培养脊的培养基并不影响脊的活性。用肢体中胚层培养 18 小时的脊制成的重组体并不像那些新鲜分离的脊那样经常产生副产物，但大多数形成的肢体在远端是完整的。这些结果表明，培养的分离的顶端外胚层脊的功能下降不仅仅是由于脊细胞死亡，而且至少部分是由于其与肢体中胚层的分离。 © 1992 Wiley-Liss, Inc.
Previously we demonstrated that high levels of insulin (5 μg/ml) permit the survival of isolated chick apical ectodermal ridge in culture (Boutin and Fallon, Dev. Biol., 104:111–116, 1984). Here we address whether lower levels of insulin or insulin‐like growth factors (IGFs) can also improve the survival of cultured apical ectodermal ridge and whether ridge function is maintained along with ridge survival. Neither IGF I nor IGF II (100 ng/ml) decreased ridge cell death; however, cell death was significantly decreased with 50 ng/ml insulin. No further improvement was obtained in the presence of both IGF I and insulin. These data suggest that insulin improved the survival of the isolated apical ectodermal ridge by binding its own receptor. To test for the maintenance of function, stage 20 ridges were cultured for 0, 6, 12, 18, or 24 hr with or without insulin (5 μg/ml or 5 ng/ml) and used to make recombinant limbs. Isolated ridges cultured for 12 hr or more produced fewer outgrowths and these were rarely distally complete. The medium in which the ridges had been cultured did not influence ridge activity, despite the major differences in cell survival. Recombinants made with ridges cultured with limb mesoderm for 18 hr did not yield outgrowths as often as those with freshly isolated ridges, but most of the limbs that did form were distally complete. These results suggest that the decline in function of cultured, isolated apical ectodermal ridge was not due merely to ridge cell death but rather, at least in part, to its separation from limb mesoderm. © 1992 Wiley‐Liss, Inc.