Effect And Significance Of TGF-β₂ On The Expression Of CD44_s In The Culture Of Human Trabecular Meshwork Cells

Purpose:To study the culture of humam trabecular meshwork cells in Vitro and observe their biological character. To investigate the effect and significance of transforming growth factor-β₂ on the expression of CD44_s in culture human trabecular meshwork cells。Methods:(1)In the first part, the trabecular meshwork tissues from human eyes were primarily Cultured. The morphologic feature and growing characteristics of the cultured cells were observed by inverted microscope, electron microscope and immunocy to chemical medthod. The proliferative curve of the cultured cells was obtained by colorimetric assay with MTT.(2) CD44_s expressed in cultured were measured by semiquantitative RT-PCR and quantitative flow cytometny. (3) CD44_s expressed in cultured Were measured by semi-quantitative RT-PCR and quantitative flow cytometry after treated with 0.0 ng/ml (comtrol),40ng/ml,80ng/ml,120ng/ml TGF-β₂ for 24h.Results:(1)The primary cells in cultured were observed from the edge of the cultured tissues after 10-15 days. The cultured cells were round , oval, fusiform and multiangular. The trabecular meshwork cells showed apical villons projections and had a high density of secondary lysosome. The junction between the trabecular meshwork cells observed most frequently were gap junction. Laminin(LN),fibronectin(FN)and neuron specific enolase (NSE) of the cultured cells expressed were identified with immunocy to chemical method. (2)The mean fluorescence in tensity of CD44_s expression of the cultured trabecular meshwork cells was 79.82±21.94,and the Value of CD44_s /G3PDH of the cells was 059±0.76.(3)The mean fluorescence intensity of CD44_s expression of the cultured trabecular cells treated was with 0.0ng/ml (cnmtrol),40ng/ml,80ng/ml,120ng/mlTGF-β₂ was 79.82±21.94,56.53±12.74,49.11±9.94 and 37.54±8.19, F=12.239, P=0.000, P＜0.001;and the Value of CD44_s/G3PDH of the cells was 0.59±0.76,0.51±0.76,0.30±0.84 and 0.06±0.04,F=135.630, P=0.000, P＜0.001, they had statistic significance. Conclusion:(1)Human trabecular meshwork cells can be successfully cultured in vitro as long as the technology of cell culture is known well. The indentification of cultured must be Cnmbined with three aspects the growing characteristic morphological features of cultured cells under light and electron microscopes and immunocy to chemical Medthod.(2)Cultured expressed CD44_s.(3) TGF-β₂ could decrease CD44_s expression in the culture of human trabecular meshwork cells. Downregulation of CD44_s expression through elevated levels TGF-β₂ might contribute to the loss of trabecular meshwork cells in the eyes of POAG.