Efficient isolation of chondrocytes from epiphyseal cartilage and their biological characteristics during culture
- VernacularTitle:原代骺板软骨细胞的高效分离和体外培养观察
- Author:
Qiang ZHOU
;
Qihong LI
;
Gang DAI
- Publication Type:Journal Article
- Keywords:
Growth plate;
Chondrocytes;
Cartilage;
Tissue engineering
- From:
Chinese Journal of Trauma
1993;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish the methods of efficiently isolating chondrocytes from epiphyseal cartilage and observe the basic biological characteristics of the cultured ones. Methods The improved isolation method through three-step enzymatic digestion using 1 g/L trypsase in 1 g/L ethylenediaminetetraacetic acid, 1 g/L hyaluronidase and 2 g/L type Ⅰ collagenase was established to isolate the primary passage chondrocytes from epiphyseal cartilage of young rabbits and its efficiency evaluated. The cellular biological characteristics of the chondrocytes were observed under the inverted and light microscopes. Results (1) All chondrocytes were isolated from the matrix after it was completely digested by three above-mentioned enzymes. The harvested chondrocytes significantly and positively correlated with the wet weight of cartilage. The number of chondrocytes from 1 g of wet cartilage was 32.3?106 with a viability of 97.6% on average. (2) The chondrocytes of the primary and the first passages showed triangle or multi-angle shapes (oval during growth and fusion) with the positive immunohistochemical stain of collagen type Ⅱ and the strong heterochromia to toluidine blue. The chondrocytes after the third passage became spindle-shaped with the negative stain of collagen type Ⅱ and the weak heterochromia to toluidine blue. Conclusions (1) The chondrocyte-isolating method through three-step enzymatic digestion has advantages of high efficiency in harvesting primary chondrocytes, high cellular viability and simple manipulation. (2) The chondrocytes differentiate and lose their special phenotypes gradually with passage.
