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10- to 12-week-old human embryonic eye globes were microdissected so that a passage was opened between the outer environment and the anterior chamber which rendered free access of tissue culture medium to the endothelial cell monolayer. The dissected eye globes were maintained in organ culture for 24 h in the continuous presence of tritiated thymidine. Cross sections were cut through the whole eye globes and subjected to autoradiographic analysis in order to estimate the mitogenic response of human embryonic corneal endothelial cells to externally supplied growth factors and hormones. It was found that the corneal endothelial cells could be stimulated to initiate DNA synthesis by exposure to insulin-like growth factor I (IGF-I). The thymidine-labelling index doubled after IGF-I supplementation. Northern blot analysis revealed the abundant presence of IGF-II transcripts in the posterior eye. In contrast, the anterior portion of the eye, including the cornea, contains barely detectable levels of IGF-II transcripts. IGF-I transcripts were detected in both parts of the eye at much lower concentrations than those for IGF-II. No insulin transcripts were found. These results demonstrate that mRNA for both IGF-I and IGF-II is present in the late first trimester eye. The observed stimulatory effects of IGF-I in organ culture suggest that local production of IGF-I and IGF-II may stimulate cell proliferation in vivo.

Stimulatory effects of insulin-like growth factors on DNA synthesis in the human embryonic cornea