2006PubMedRequires access

[Primary culture of human pterygia cells in vitro].

Shangchen Hao, Zuguo Liu

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Abstract

PURPOSE: To probe a selective method of isolate and culture of human pterygia cells in vitro. METHODS: Fifty primary pterygia were got by operation and cut to clips less than 1 x 1 mm2, segments were digested by collagenase II for 20 minutes and by trypsin for 10 minutes under 37 degrees C, and then centrifugated for 15 minutes by 1500 rpm. The collected cells were cultured in conditioned medium for endothelial cells. After purified by scrape and density gradient centrifugation, the cells were detected and identified by immunohistochemistry and electron microscope. RESULTS: The purity of cultured fibroblasts and endothelial cell type cells were more than 90%. CD31, CD34, and Weibel-Palade bodies were performed in these cells cytoplasm. CONCLUSION: We obtained the method of culturing human pterygia fibroblasts and endothelial cell type cells and it is a feasible and convenient approach.

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PURPOSE: To probe a selective method of isolate and culture of human pterygia cells in vitro. METHODS: Fifty primary pterygia were got by operation and cut to clips less than 1 x 1 mm2, segments were digested by collagenase II for 20 minutes and by trypsin for 10 minutes under 37 degrees C, and then centrifugated for 15 minutes by 1500 rpm. The collected cells were cultured in conditioned medium for endothelial cells. After purified by scrape and density gradient centrifugation, the cells were detected and identified by immunohistochemistry and electron microscope. RESULTS: The purity of cultured fibroblasts and endothelial cell type cells were more than 90%. CD31, CD34, and Weibel-Palade bodies were performed in these cells cytoplasm. CONCLUSION: We obtained the method of culturing human pterygia fibroblasts and endothelial cell type cells and it is a feasible and convenient approach.

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Available abstract

PURPOSE: To probe a selective method of isolate and culture of human pterygia cells in vitro. METHODS: Fifty primary pterygia were got by operation and cut to clips less than 1 x 1 mm2, segments were digested by collagenase II for 20 minutes and by trypsin for 10 minutes under 37 degrees C, and then centrifugated for 15 minutes by 1500 rpm. The collected cells were cultured in conditioned medium for endothelial cells. After purified by scrape and density gradient centrifugation, the cells were detected and identified by immunohistochemistry and electron microscope. RESULTS: The purity of cultured fibroblasts and endothelial cell type cells were more than 90%. CD31, CD34, and Weibel-Palade bodies were performed in these cells cytoplasm. CONCLUSION: We obtained the method of culturing human pterygia fibroblasts and endothelial cell type cells and it is a feasible and convenient approach.

Key concepts: Collagenase, CD31, In vitro, Primary culture, Cell culture, Cytoplasm, Molecular biology, Differential centrifugation

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