In Vitro Culture and Biological Characterization of Meniscal Fibrochondrocytes.

Abstract

Background: Currently, there is a lot of discussion in clinical and scientific research over the best ways to delay meniscal degeneration and speed up its healing. Meniscal injury and degeneration are significant contributors to the development of knee osteoarthritis in a pathological state.

Objective: To isolate, culture and characterize rat meniscal fibrochondrocytes in vitro, and to provide a simple and feasible cell culture method for the study of damage repair of rat meniscal fibrochondrocytes.

Methods: The rat medial and lateral meniscus of both knees was surgically isolated. Trypsin and type II collagenase were used to remove the cells, and toluidine blue staining and type II collagen immunofluorescence were used to identify the cells. The cells were then routinely cultured in low-sugar DMEM complete culture medium.

Results: At different time points, cells showed different physiological shapes, from polygonal or short spindle to spindle shape, and finally to triangle or ellipse, and cell proliferation ability gradually increased with time. The OD values of cells cultured at 48h and 72h were higher than those at 24h. Comparing OD values of cells cultured for 48h and 72h, although OD value of 72h increased. Toluidine blue staining and type I collagen immunofluorescence staining were positive.

Conclusion: A more dependable technique for fibrochondrocyte isolation and culture is offered for the study of meniscus in molecular biology and tissue engineering. The cells cultured using this method are morphologically stable, have a strong proliferation ability, and possess the fundamental biological properties of fibrochondrocytes in vivo.