[Human trabeculer cells in culture and cell identification].
OBJECTIVE
To establish the method of culturing human trabecular cells(HTCs) in vitro and to determine identifyng main points concerning cultured HTCs.


METHODS
Trabecular meshwork, corneal and scleral tissue were collected from sixty-eight eyeballs of thirty-four human donors less than six years of age within 24 hours after death. The tissues were primarily cultured and the cells subcultured. Cultured cells were observed by light and electron microscopes. Fibronectin(FN), laminin(LN) and IV collagen (IV C) in extracellular matrix(ECM) of the cultured cells were immunohistocytochemically stained by S-P method.


RESULTS
HTCs easily grew in the event of young donors and short-time drawing materials. Trabecular meshwork should be accurately separated. Techniques in primary culture, and time and proportion in subculture should be suitabled. Experimental studies should select the third to fifth passages. The growing characteristics, morphological features of cultured HTCs under light and electron microscopes differed from those of corneal and scleral cells adjoining them. Ultramicrostucture distinguished HTCs from corneal interstitial cells. ECM stained by S-P method distinguished HTCs from scleral fibroblast.


CONCLUSIONS
These studies suggest that it is not difficult to culture HTCs in general culturing condition as long as the main points about culturing them are known-well. The identification of cultured HTCs must be combined with three aspects: the growing characteristics and morphological features of cultured cells under light and electron microscopes, and the immunohistochemical stained peculiarities of FN, LN and IV C in ECM of cultured cells. To establish the method of culturing HTCs in vitro and to determine identifyng main points concerning cultured HTCs become an essential condition and an important guarantee in order to investigate the pathogenesis and mechanism on primary open angle glaucoma(POAG) deeply.
Zhong
L
L
Department of Ophthalmology, First School of Clinical Medicine, Beijing Medical University, Beijing 100034, China.
Li
M
M
Li
M
M
D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003315:Cornea / Q000166:cytology; D005109:Extracellular Matrix / Q000737:chemistry*; D005353:Fibronectins / Q000032:analysis; D005902:Glaucoma, Open-Angle / Q000209:etiology; D006801:Humans; D007150:Immunohistochemistry; D007223:Infant; D007797:Laminin / Q000032:analysis; D012590:Sclera / Q000166:cytology; D014129:Trabecular Meshwork / Q000166:cytology* / Q000378:metabolism
D005353:Fibronectins; D007797:Laminin