[Isolation and gene screening for collagen phagocytic subpopulation of fibroblasts and non-collagen phagocytic subpopulation of fibroblasts].

Objective: To isolate the collagen phagocytic subpopulation of fibroblast(CPSF) and noncollagen phagocytic subpopulation of fibroblast(n CPSF) and to identify their differentially expressed genes.Methods: The CPSF and n CPSF was isolated by using collagen-fluorescein-isothiocynate-latex bead(COL-FITC-LB) phagocytosis technique and FCM sorting method. Microarray analysis was used to screen the differentially expressed genes, which were verified by real-time PCR. Results: CPSF and n CPSF was successfully isolated. Seventeen differentially expressed genes were identified. Compared with nC PSF, the expression of 12 or 5 genes was up-regulated or downregulated in CPSF. Three of the 12 up-regulated genes were urokinase plasminogen activator receptor-associated protein(u PARAP), cytochrome b-245, beta polypeptide(CYBB) and Hook homolog 1(HOOK1), which were confirmed by real-time PCR. u PARAP m RNA expression level in CPSF was 2788 times of that in nC PSF. CYBB m RNA expression in CPSF was only 0.85 times of that in nC PSF. HOOK1 m RNA expression in CPSF was 1.96 times of that in nC PSF(P0.05). Conclusion: A novel method is successfully established to isolate CPSF and n CPSF. u PARAP is the main differentially expressed gene in CPSF and n CPSF, which is obviously involved in the fibroblast collagen phagocytosis. It might be a potential biomarker for treatment of collagen diseases.