Crow J M, Lima P H, Agapitos P J, Nelson J D
Department of Ophthalmology, Ramsey Clinic, St. Paul, MN.
Invest Ophthalmol Vis Sci. 1994 Jan;35(1):128-33.
To investigate the effects of insulin, epidermal growth factor (EGF), and the corneal storage media--DexSol--at 24 and 48 hours on DNA synthesis in confluent primary cultures of bovine corneal endothelial cells.
Flow cytometry was used to measure changes in DNA synthesis. This technique allows a large number of cells to be counted and sorted into G1, S, and G2/M phases of the cell cycle.
Changing the normal culture media to DexSol had no effect on the cell cycle at 24 or 48 hours. The addition of insulin, EGF, or insulin + EGF to DexSol increased DNA synthesis within 24 hours. The mitotic indices for DexSol, DexSol + insulin, and DexSol + EGF were 0.134 (SE = +/- 0.022), 0.207 (+/- 0.027), and 0.205 (+/- 0.052), respectively. Adding insulin + EGF to the DexSol resulted in the most significant change in S and G2/M, increasing the mitotic index to 0.300 (+/- 0.072) (P = 0.0116). At 48 hours, the presence of the growth factors no longer had any effect.
Flow cytometry was a useful technique in separating cultured bovine corneal endothelial cells according to their DNA content. Analysis of the cultures after the addition of insulin and EGF showed an increase in DNA synthesis. The synergistic effects of the growth factors on corneal endothelial cells suggest that they stimulate mitotic activity by different mechanisms. The addition of mitogens to eye bank storage media may increase corneal endothelial cell densities in donor corneas.
研究胰岛素、表皮生长因子（EGF）以及角膜储存介质——DexSol——在24小时和48小时时对牛角膜内皮细胞汇合原代培养物中DNA合成的影响。
采用流式细胞术测量DNA合成的变化。该技术可对大量细胞进行计数并将其分选到细胞周期的G1、S和G2/M期。
将正常培养基更换为DexSol在24小时或48小时时对细胞周期无影响。向DexSol中添加胰岛素、EGF或胰岛素 + EGF可在24小时内增加DNA合成。DexSol、DexSol + 胰岛素和DexSol + EGF的有丝分裂指数分别为0.134（标准误 = ±0.022）、0.207（±0.027）和0.205（±0.052）。向DexSol中添加胰岛素 + EGF导致S期和G2/M期变化最为显著，有丝分裂指数增加至0.300（±0.072）（P = 0.0116）。在48小时时，生长因子的存在不再有任何影响。
流式细胞术是一种根据DNA含量分离培养的牛角膜内皮细胞的有用技术。添加胰岛素和EGF后对培养物的分析显示DNA合成增加。生长因子对角膜内皮细胞的协同作用表明它们通过不同机制刺激有丝分裂活性。向眼库储存介质中添加促有丝分裂剂可能会增加供体角膜中角膜内皮细胞的密度。
