Study of cryopreservation of articular chondrocytes using the Taguchi method.
This study evaluates the effect of control factors on cryopreservation of articular cartilage chondrocytes using the Taguchi method. Freeze-thaw experiments based on the L(8)(2(7)) two-level orthogonal array of the Taguchi method are conducted, and ANOVA (analysis of variables) is adopted to determine the statistically significant control factors that affect the viability of the cell. Results show that the type of cryoprotectant, freezing rate, thawing rate, and concentration of cryoprotectant (listed in the order of influence) are the statistically significant control factors that affect the post-thaw viability. The end temperature and durations of the first and second stages of freezing do not affect the post-thaw viability. Within the ranges of the control factors studied in this work, the optimal test condition is found to be a freezing rate of 0.61+/-0.03 degrees C/min, a thawing rate of 126.84+/-5.57 degrees C/min, Me(2)SO cryoprotectant, and a cryoprotectant concentration of 10% (v/v) for maximum cell viability. In addition, this study also explores the effect of cryopreservation on the expression of type II collagen using immunocytochemical staining and digital image processing. The results show that the ability of cryopreserved chondrocytes to express type II collagen is reduced within the first five days of monolayer culture.
Buddhist Tzu-Chi Dalin General Hospital, Chiayi, Taiwan.
Shaw-Ruey
SR
Lyu
Wei Te
WT
Wu
Chien Chih
CC
Hou
Wen-Hsin
WH
Hsieh
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology / Q000378:metabolism; D002470:Cell Survival; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism; D024043:Collagen Type II / Q000378:metabolism; D015925:Cryopreservation / Q000379:methods*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D013552:Swine; D013997:Time Factors
D024043:Collagen Type II