Lu L, Garcia C A, Mikos A G
Department of Chemical Engineering and Institute of Biosciences and Bioengineering, Rice University, Houston, TX 77251-1892, USA.
J Biomater Sci Polym Ed. 1998;9(11):1187-205. doi: 10.1163/156856298x00721.
Thin films of 50:50 and 75:25 poly(DL-lactic-co-glycolic acid) (PLGA) were manufactured with a controlled thickness of less than 10 microm. The effect of PLGA copolymer ratio on in vitro cell attachment, proliferation, morphology, and tight junction formation was evaluated using a human D407 retinal pigment epithelium (RPE) cell line. Almost complete cell attachment was achieved on both PLGA films after 8 h of cell seeding, which was comparable to that on tissue culture polystyrene (TCPS) controls. The initial cell seeding density affected attachment, and the optimal value for 50:50 PLGA was 25000 cells cm(-2). After 7 days of in vitro culture, cell density on 50:50 and 75:25 PLGA films increased 45 and 40 folds, respectively, and a 34-fold increase was observed on TCPS. The RPE cells cultured on PLGA films at confluence had a characteristic cobblestone morphology. Confluent RPE cells also developed normal tight junctions in vitro which were concentrated mainly at the apical surfaces of cell-cell junctions. These results demonstrated that thin biodegradable PLGA films can provide suitable substrates for human RPE cell culture, and may serve as temporary carriers for subretinal implantation of organized sheets of RPE.
制备了50:50和75:25的聚（DL-乳酸-乙醇酸共聚物）（PLGA）薄膜，其控制厚度小于10微米。使用人D407视网膜色素上皮（RPE）细胞系评估了PLGA共聚物比例对体外细胞附着、增殖、形态和紧密连接形成的影响。细胞接种8小时后，两种PLGA薄膜上几乎都实现了完全细胞附着，这与组织培养聚苯乙烯（TCPS）对照相当。初始细胞接种密度影响附着，50:50 PLGA的最佳值为25000个细胞/cm²。体外培养7天后，50:50和75:25 PLGA薄膜上的细胞密度分别增加了45倍和40倍，TCPS上观察到增加了34倍。在PLGA薄膜上汇合培养的RPE细胞具有特征性的鹅卵石形态。汇合的RPE细胞在体外也形成了正常的紧密连接，主要集中在细胞-细胞连接的顶端表面。这些结果表明，可生物降解的PLGA薄膜可为人类RPE细胞培养提供合适的基质，并可作为RPE有组织薄片视网膜下植入的临时载体。
