title: Differentiation of bone marrow-derived mesenchymal stem cells into chondrocytes             using chondrocyte extract.
abstract: Reprogramming cells using cell extracts is an effective method for harvesting cells of interest. The aim of this study was to investigate the chondrogenic transdifferentiation potential of swine bone marrow-derived mesenchymal stem cells (BM-MSCs) by culturing with chondrocyte extract, using monolayer and micromass culture. Chondrogenic-specific markers were detected via reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence. After 7 days of induction in monolayer culture, BM-MSCs reversibly permeabilized with streptolysin O (SLO), a bacterial exotoxin that is capable of forming large pores in the plasma membrane of mammalian cells, and expressed chondrocyte‑specific genes such as type II collagen (COL II) and aggrecan. A positive protein expression of COL II was also observed. However, BM-MSCs treated without SLO did not express the related genes and proteins. The transition of reprogrammed BM-MSCs was lost 14 days later. By using micromass culture, reprogrammed BM-MSCs were able to maintain the change until the 14th day. In summary, permeabilized BM-MSCs were transiently transdifferentiated into chondrocytes by co-culturing with the chondrocyte extract. Moreover, a high-density culture method was able to increase the time in which the phenotypic change was maintained.
authors: Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai, PR China.
authors: Xinjie
authors: X
authors: Tang
authors: Lingling
authors: L
authors: Sheng
authors: Feng
authors: F
authors: Xie
authors: Qun
authors: Q
authors: Zhang
mesh_terms: D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001426:Bacterial Proteins / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000166:cytology*; D002463:Cell Membrane Permeability / Q000187:drug effects; D054338:Cell Transdifferentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D018920:Coculture Techniques; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000473:pathology; D013301:Streptolysins / Q000494:pharmacology; D013552:Swine
chemical_list: D053674:Aggrecans; D001426:Bacterial Proteins; D024043:Collagen Type II; D013301:Streptolysins; C025964:streptolysin O