Immortalization and characterization of mouse temporomandibular joint disc cell clones with capacity for multi-lineage differentiation

Abstract

Objective Despite the importance of TMJ disc in normal function and disease, studying the responses of its cells has been complicated by the lack of adequate characterization of the cell subtypes. The purpose of our investigation was to immortalize, clone, characterize and determine the multi-lineage potential of mouse TMJ disc cells. Design Cells from 12-week-old female mice were cultured and immortalized by stable transfection with human telomerase reverse transcriptase. The immortalized cell clones were phenotyped for fibroblast- or chondrocyte-like characteristics and ability to undergo adipocytic, osteoblastic and chondrocytic differentiation. Results Of 36 isolated clones, four demonstrated successful immortalization and maintenance of stable protein expression for up to 50 passages. Two clones each were initially characterized as fibroblast-like and chondrocyte-like on the basis of cell morphology and growth rate. Further the chondrocyte-like clones had higher mRNA expression levels of cartilage oligomeric matrix protein (>3.5-fold), collagen × (>11-fold), collagen II expression (2-fold) and collagen II:I ratio than the fibroblast-like clones. In contrast, the fibroblast-like clones had higher mRNA expression level of vimentin (>1.5-fold), and fibroblastic specific protein 1 (>2.5-fold) than he chondrocyte-like clones. Both cell types retained multi-lineage potential as demonstrated by their capacity to undergo robust adipogenic, osteogenic and chondrogenic differentiation. Conclusions These studies are the first to immortalize TMJ disc cells and characterize chondrocyte-like and fibroblast-like clones with retained multi-differentiation potential that would be a valuable resource in studies to dissect the behavior of specific cell types in health and disease and for tissue engineering.

Conditional Deletion of Fgfr3 in Chondrocytes leads to Osteoarthritis-like Defects in Temporomandibular Joint of Adult Mice

“…The collagen II is also expressed in TMJ disc cells ( Fig. 1B ), which is in consistent with previous studies 29 30 . We found that the expression of FGFR3 in TMJ disc cells was significantly down-regulated in Fgfr3 cKO mice ( Fig.…”

“…Additionally, the TMJ disc, a specialized fibrocartilaginous tissue, is similar to the meniscus in knee joints. Both of the chondrocyte- and fibroblast-like cell clones was detected in the TMJ disc 29 . The collagen II is also expressed in TMJ disc cells ( Fig.…”

Efficient and simple approach to in vitro culture of primary epithelial cancer cells

“…Obviously, the last approach is associated with the risk of altering molecular profile of the cell and hindering the precise reflection of in vivo state [35,36]. Nevertheless, as our approach did not prevent the inevitable inhibition of in vitro cancer cells proliferation, we made an attempt to transduce primary cancer cells with well-known immortalizing genes– SV40LT [37], BMI-1 [38] and hEST2 [39]. None of the utilized genes significantly prolonged culture time, neither in breast nor in prostate cells.…”

Effects of dynamic radial tensile stress on fibrocartilage differentiation of bone marrow mesenchymal stem cells

“…Approximately 63-70% of the cells in the mature TMJ disc are the elongated spindle shaped fibroblast-like cells (called fibrochondrocytes) [5,26], and these cells primarily expressed the fibroblast-specific protein 1 (FSP1) [26]. Immunofluorescence staining revealed that the expression of fibrochondrocyte marker FSP1 appeared in the stretched BMSCs, but not in the control group ( Fig.…”