Chondrogenesis from immortalized human mesenchymal stem cells: comparison between collagen gel and pellet culture methods.
Human mesenchymal stem cells (hMSCs) can differentiate into cells of connective tissue lineages, including cartilage. To overcome the limiting autogenous chondrocyte populations available in cartilage repair, various methods have been developed to maximize chondrogenesis of hMSCs in vitro, most of which use cells derived from primary culture. In this study, we compared chondrogenesis of immortalized hMSCs embedded in collagen gel to those grown in pellet culture. The hMSCs in collagen scaffolds expressed more glycosaminoglycan than those in pellet culture. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) analysis demonstrated that the expression of genes encoding sox-9, aggrecan, and types I and II collagen increased in pellet culture over time. However, in the collagen cultures, only type II collagen and aggrecan expression increased over time, whereas sox-9 expression remained unchanged and type I collagen expression decreased. These results indicate that the immortalized hMSC line is a promising tool for further in vitro chondrogenic studies.
Chang
Chih-Hung
CH
Department of Surgery, Division of Orthopedics, Far Eastern Memorial Hospital, Taipei, Taiwan.
Lin
Hsin-Yi
HY
Fang
Hsu-Wei
HW
Loo
Siow-Tung
ST
Hung
Shih-Chieh
SC
Ho
Yi-Ching
YC
Chen
Chia-Chun
CC
Lin
Feng-Huei
FH
Liu
Hwa-Chang
HC
D053674:Aggrecans / Q000235:genetics; D018929:Cell Culture Techniques; D002460:Cell Line; D020219:Chondrogenesis / Q000502:physiology*; D024042:Collagen Type I / Q000235:genetics; D024043:Collagen Type II / Q000235:genetics*; D004247:DNA / Q000032:analysis; D015870:Gene Expression; D006025:Glycosaminoglycans / Q000235:genetics; D006609:High Mobility Group Proteins / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055758:SOX9 Transcription Factor; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds; D014157:Transcription Factors / Q000235:genetics
D053674:Aggrecans; D024042:Collagen Type I; D024043:Collagen Type II; D006025:Glycosaminoglycans; D006609:High Mobility Group Proteins; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D014157:Transcription Factors; D004247:DNA