Maintenance of human chondrogenic phenotype on a dendrimer-immobilized surface for an application of cell sheet engineering

BMC Biotechnology

Sopita Wongin¹  Kwanchanok Viravaidya-Pasuwat¹  Mee-Hae Kim²  Masahiro Kino-oka²  Saranatra Waikakul³  Pojchong Chotiyarnwong³  Wanwipa Siriwatwechakul⁴ 

[1] Biological Engineering Program, Faculty of Engineering, King Mongkut’s University of Technology Thonburi;Department of Biotechnology, Graduate School of Engineering, Osaka University;Department of Orthopaedic Surgery, Faculty of Medicine, Siriraj Hospital, Mahidol University;School of Bio-Chemical Engineering and Technology, Sirindhorn International Institute of Technology, Thammasat University;

关键词: Human chondrocytes;    Dendrimer surface;    Morphological change;    Chondrocyte sheet;    Stress fiber formation;    Extracellular matrix formation;   

DOI  :  10.1186/s12896-018-0426-1

来源: DOAJ

【 摘 要 】

Abstract Background Dedifferentiation of chondrocytes during cell expansion is one of the barriers in tissue construction for cartilage repair. To understand chondrocyte behavior and improve cell expansion in monolayer culture, this study investigated the effects of morphological changes and cellular aggregation on the maintenance of chondrogenic capacity by observing the expression patterns of chondrogenic (collagen type II and aggrecan) and dedifferentiation (collagen type I) markers. Primary human chondrocytes were cultured on either a polystyrene surface (PS) or a polyamidoamine dendrimer surface with a fifth-generation (G5) dendron structure to create a one-step process of cell expansion and the maintenance of chondrogenic activities prior to the construction of cell sheets. Results During the first two passages (P0 - P2), the relative mRNA level of collagen type II decreased in all cultures, while that of collagen type I increased. Remarkably, the level of collagen type II was higher and aggrecan was retained in the chondrocytes, forming cell aggregates and showing some round-shaped cells with less production of stress fibers on the G5 surface compared to fibroblast-like chondrocytes with abundant stress fibers on the PS surface. The numbers of P2 chondrocytes on the G5 and PS surfaces were nearly the same and sufficient for construction of chondrocyte sheets using a temperature-responsive plate. Without a supporting material during cell sheet manipulation, chondrocyte sheets spontaneously detached and exhibited a honeycomb-like structure of stress fibers. Unlike the chondrocyte sheets constructed from cells on the PS surface, the chondrocyte sheets from cells on the G5 surface had higher chondrogenic activities, as evidenced by the high expression of chondrogenic markers and the low expression of dedifferentiation markers. Conclusions The one-step process of cell expansion and maintenance of chondrogenic activity could be obtained using the G5 surface. Human chondrocyte sheets were successfully constructed with high chondrogenic activity. These findings may lead to an alternative cultivation technique for human chondrocytes that offers high clinical potential in autologous chondrocyte implantation.

【 授权许可】

Unknown