Differentiation of bone marrow-derived mesenchymal stem cells into chondrocytes using chondrocyte extract

Abstract

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 highdensity culture method was able to increase the time in which the phenotypic change was maintained.

Reprogramming human adipose tissue stem cells using epidermal keratinocyte extracts

“…In recent years, there have been significant advancements in tissue-engineered skin replacements ( 1 – 3 ); however, a good source of autogeneic cells remains to be determined. Nuclear reprogramming offers a promising method, whereby extracts from a specific type of cell are introduced into donor cells and the donor cells subsequently acquire the characteristics of the cells from which the extracts were derived ( 4 ). This approach consists of incubating reversibly permeabilized donor cells in nuclear and cytoplasmic extracts derived from another type of somatic cell.…”

The role of mechanobiology in bone and cartilage model systems in characterizing initiation and progression of osteoarthritis

“…Such systems have been employed to determine, for example, that co-culturewith chondrocytes can improve the chondrogenic differentiation of MSCs through mechanisms involving both soluble factor secretion and cell-cell contact. [224][225][226] As with monolayer cultures, combining these co-cultures with methods to mechanically stimulate the cells will provide new insights into their function and behavior. For example, combining tensile stimulation with co-cultures of MSCs and chondrocytes was reported to increase chondrogenic phenotypes and rapidity of cell expansion.…”