Agar, meets USP testing specifications

Agar meeting USP testing specifications is used for clonal culture, phenotypic identification, purification and screening, and medium performance evaluation of chick embryo chondrocytes^[2].
1. Preparation of soft agar culture system
Unpurified Difco bacterial agar and tryptone phosphate are used to prepare the basal agar layer. Diluted single-cell suspension (10³ cells/60 mm culture dish) is inoculated on the upper layer. Modified Eagle's medium or F-12 medium containing 10% fetal bovine serum serves as the culture medium, and aseptic operation is performed throughout the process.
2. Results of clonal culture and phenotypic maintenance of chondrocytes
(1) After inoculation, chondrocytes derived from tibiae/femora of 13-day-old chick embryos initiate division and secrete chondromucoprotein matrix within several days, and form colonies containing more than 1000 cells in 3 weeks;
(2) Chondrocyte clones isolated from agar (designated as Ag series), after being passaged to monolayer culture, can still form colonies and produce metachromatic matrix when re-inoculated into soft agar, thus stably maintaining the differentiated phenotype of chondrocytes;
(3) Agarose can replace agar to achieve the same chondrocyte colony-forming effect.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.