Abstract
Recent advancement in cartilage tissue engineering has explored the potential of 3D culture to mimic the in vivo environment of human cartilaginous tissue. Three-dimensional culture using microspheres was described to play a role in driving the differentiation of mesenchymal stem cells to chondrocyte lineage. However, factors such as mechanical agitation on cell chondrogenesis during culture on the microspheres has yet to be elucidated. In this study, we compared the 2D and 3D culture of bone-marrow-derived mesenchymal stem cells (BMSCs) on gelatin microspheres (GMs) in terms of MSC stemness properties, immune-phenotype, multilineage differentiation properties, and proliferation rate. Then, to study the effect of mechanical agitation on chondrogenic differentiation in 3D culture, we cultured BMSCs on GM (BMSCs-GM) in either static or dynamic bioreactor system with two different mediums, i.e., F12: DMEM (1:1) + 10% FBS (FD) and chondrogenic induction medium (CIM). Our results show that BMSCs attached to the GM surface and remained viable in 3D culture. BMSCs-GM proliferated faster and displayed higher stemness properties than BMSCs on a tissue culture plate (BMSCs-TCP). GMs also enhanced the efficiency of in-vitro chondrogenesis of BMSCs, especially in a dynamic culture with higher cell proliferation, RNA expression, and protein expression compared to that in a static culture. To conclude, our results indicate that the 3D culture of BMSCs on gelatin microsphere was superior to 2D culture on a standard tissue culture plate. Furthermore, culturing BMSCs on GM in dynamic culture conditions enhanced their chondrogenic differentiation.
Highlights
Tissue Engineering and regenerative medicine have been considered as a potential alternative to conventional treatment for cartilage diseases [1]
Our current work aims to explore the potential effect of the 3D culture system in a dynamic culture environment on gelatin microspheres (GMs) cultured with bone-marrow-derived Mesenchymal stem cells (MSCs) (BMSCs)
This study demonstrated the influence of 3D culture conditions, compared to the 2D monolayer culture, on the proliferation and chondrogenic differentiation of bone-marrow-derived mesenchymal stem cells (BMSCs)
Summary
Tissue Engineering and regenerative medicine have been considered as a potential alternative to conventional treatment for cartilage diseases [1]. Numerous preclinical studies have shown stem cells’ efficacy on reviving damaged cartilage tissues to its functional state [2,3,4]. More than 200 clinical trials have been conducted to treat cartilage damage using stem cells to determine the safety, efficacy, and long-term feasibility in patients [5], some of these were proven effective, while some were not [5]. It was reported that MSCs would tend to lose their chondrogenic potential with culturing time [11]. This claim could explain the failures of several clinical studies beforehand. There is a necessity to fill this gap in order to preserve its functionality and regenerative capabilities
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