Ibaraki N, Lin L R, Reddy V N
Eye Research Institute, Oakland University, Rochester, MI 48309-4401, USA.
Invest Ophthalmol Vis Sci. 1995 Oct;36(11):2304-12.
A successful method to subculture human lens epithelial (HLE) cells that retain their intrinsic characteristics is of great importance. This study examines the effects of four different growth factors on proliferation and differentiation in HLE cells in early subcultures.
Specimens of HLE cells were obtained from infants. First- or second-passage cells were cultured in the presence of 10(-2) to 10(2) ng/ml acidic and basic fibroblast growth factor (aFGF, bFGF), epidermal growth factor (EGF), or insulin-like growth factor-I (IGF-I). Cell proliferation was determined from cell number, and fiber differentiation was assessed from the time of appearance, the number of lentoids formed, and the expression of gamma-crystallin.
Cell proliferation was increased by EGF, bFGF, and IGF-I at concentrations greater than 10(-1) ng/ml; the most effective concentration was 10 ng/ml. The effect of aFGF on proliferation appeared only at a concentration of 10(2) ng/ml. EGF, bFGF, or IGF-I at 10 ng/ml affected the time of appearance and the number of lentoids formed within 5 to 7 days. In contrast, lentoids were observed after 42 days without the addition of growth factors. Lentoid formation was accompanied by the expression of gamma-crystallin.
EGF, aFGF, bFGF, and IGF-I stimulated cell proliferation and fiber differentiation in early subcultures.
建立一种能成功传代培养并保持其固有特性的人晶状体上皮（HLE）细胞的方法至关重要。本研究探讨了四种不同生长因子对早期传代培养的HLE细胞增殖和分化的影响。
从婴儿获取HLE细胞标本。将第一代或第二代传代细胞在含有10⁻²至10² ng/ml酸性和碱性成纤维细胞生长因子（aFGF、bFGF）、表皮生长因子（EGF）或胰岛素样生长因子-I（IGF-I）的条件下培养。通过细胞数量测定细胞增殖情况，并从出现时间、形成的类晶状体数量以及γ-晶状体蛋白的表达来评估纤维分化情况。
浓度大于10⁻¹ ng/ml时，EGF、bFGF和IGF-I可促进细胞增殖；最有效浓度为10 ng/ml。aFGF仅在浓度为10² ng/ml时对增殖有影响。10 ng/ml的EGF、bFGF或IGF-I在5至7天内影响类晶状体的出现时间和形成数量。相比之下，不添加生长因子时，42天后才观察到类晶状体。类晶状体形成伴随着γ-晶状体蛋白的表达。
EGF、aFGF、bFGF和IGF-I在早期传代培养中可刺激细胞增殖和纤维分化。
