Iwig M, Glässer D, Luther M, Struck H G
Institut für Physiologische Chemie, Medizinische Fakultät, Martin-Luther-Universität Halle-Wittenberg, Hollystr. 1, 06097 Halle.
Klin Monbl Augenheilkd. 2001 Feb;218(2):102-10. doi: 10.1055/s-2001-12253.
Bovine lens epithelial cells in culture revealed a high sensitivity against micromolar concentrations of linoleic acid. To prove the assumption that unsaturated free fatty acids are risk factors for cataractogenesis, human lens cell lines are needed. Furthermore, the reactivation of nucleus-containing fiber cells to mitotic growth may hint at their role in after cataract genesis.
Epithelium-capsule-preparations obtained by capsulorhexis were cultured in serum containing medium. Subculturing of these adult human lens epithelial cells was done by trypsinization. Fiber cell bundles from the equator region of a fetal human lens were transferred into culture medium. Aggregates of nucleus containing fiber cells were isolated from floating fiber cell bundles by trypsinization. Subculturing and cryoconservation of suitable cell lines.
Primary culture of epithelium-capsule-preparations results in flattening, migration and proliferation of adult human lens epithelial cells. Nucleus containing fiber cells were reactivated to mitotic growth after adhesion to a suitable substratum. Established cell lines were received from adult human lens epithelial cells and fetal human fiber cells after repeated subculturing.
Lens-capsule-preparations available from cataract surgery are well suited for the isolation of human lens cell lines, which were needed for testing cytotoxicity of drugs and for tracing of cataractogenic risk factors. The finding that nucleus containing fiber cells from the equator of human lenses can be reactivated to proliferating cells let us suppose, that these cells, which can not be removed easily from the posterior lens capsule, contribute to the after cataract formation.
培养的牛晶状体上皮细胞对微摩尔浓度的亚油酸表现出高敏感性。为了证明不饱和游离脂肪酸是白内障发生的危险因素这一假设,需要人类晶状体细胞系。此外,含核纤维细胞重新激活至有丝分裂生长可能暗示它们在后发性白内障发生中的作用。
通过撕囊术获得的上皮 - 囊膜制剂在含血清培养基中培养。这些成人人类晶状体上皮细胞的传代培养通过胰蛋白酶消化进行。将来自人类胎儿晶状体赤道区域的纤维细胞束转移至培养基中。通过胰蛋白酶消化从漂浮的纤维细胞束中分离出含核纤维细胞聚集体。对合适的细胞系进行传代培养和冷冻保存。
上皮 - 囊膜制剂的原代培养导致成人人类晶状体上皮细胞扁平、迁移和增殖。含核纤维细胞在黏附于合适的基质后重新激活至有丝分裂生长。经过反复传代培养后,从成人人类晶状体上皮细胞和胎儿人类纤维细胞中获得了已建立的细胞系。
白内障手术中可获得的晶状体囊膜制剂非常适合用于分离人类晶状体细胞系,这对于测试药物的细胞毒性和追踪白内障发生风险因素是必需的。人类晶状体赤道部含核纤维细胞可重新激活为增殖细胞这一发现使我们推测,这些不易从晶状体后囊膜清除的细胞会导致后发性白内障的形成。