Observation On Isolation Of Rabbit Articular Chondrocytes By Collagenase Digestion And In Vitro Aggregation Culture
Keywords
Chondrocytes, Collagenase, Aggregate Culture, Articular Cartilage, Rabbit
Abstract
Objective To confirm the reliability of this acquisition method by observing the biological properties of rabbit articular chondrocytes obtained by digestion and isolation with a single collagenase and aggregation and culture in vitro. Methods: Articular cartilage from living adult rabbits was collected, digested with type II collagenase in vitro, and then cultured in high-density aggregation to observe cell morphology and growth using an inverted microscope. Phenotypic changes of chondrocytes were observed with conventional staining and type II collagen immunohistochemical staining. . Results (1) The total number of collected cells is (1.2~2.6)¡Á105? g-1. The average cell viability rate detected by trypan blue was 96.7%. (2) The first three generations of cells grow rapidly and take on the shapes of short spindles, triangles, polygons, etc., and become oval when fused. (3) After aggregation amplification and culture, Mallory and toluidine blue metachromatic reactions were both positive, and type II collagen immunohistochemistry results were positive. Conclusion A large number of excellent chondrocytes can be obtained by digesting and separating with a single collagenase and aggregating and culturing them in vitro.
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Original research was done by Ma Tao, Yang Xiaoyu, Wen Peng, Wan Jun, Luo Xiaojun
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