Biopsies removed from 57 patients considered for cartilage transplantation
were grown at CTI Ltd. (47 biopsies) and at Tel Aviv University (10 biopsie
s). Tissue processing took place in dedicated laboratories. Explant culture
s allowed cell number expansion. Fifty-four out of 57 biopsies grew cells.
Fanning out of the cells began after 5-15 days in culture. Two passages lat
er, cell numbers in the 10(7) range were achieved. Cells from all cultures
expressed mRNA of aggrecan and link protein but not of alkaline phosphatase
. Histochemical stains such as alcian blue pH 1 were negative in sparse mon
olayer cultures, but positive in pellet cultures. Immunohistochemistry demo
nstrated expression of collagen type I in monolayer cultures, switching to
collagen type II in micromass cultures. Fibroblast growth factor receptor 3
, a recently described characteristic receptor of precartilaginous cells, w
as expressed in monolayers and disappeared in micromass cultures. In conclu
sion, explants of articular chondrocytes cultured in vitro consistently yie
ld monolayer cultures. The cells appear to revert to dedifferentiated chond
rocytes, expressing a mesenchymal stem cell protein profile. Simultaneously
, these cells regained their capacity to proliferate. Cultures held as micr
omass allowed reexpression of the differentiated phenotype traits.