Immortalization of Human Cartilage Endplate Stem Cell from Degenerated Human Intervertebral Disc

Abstract

Introduction Low back pain is one of the most common reasons for seeking medical advice and is a major cause of work-related disabilities. Recently, cell-based therapies for regenerating or repairing the degenerated disc have become treatment options. Our group previously demonstrated that stem cells exit in human degenerated CEP and that cartilage endplate-derived stem cells (CESCs) are superior to BM-MSCs in terms of osteogenesis and chondrogenesis. CESCs may represent one of the most promising stem cells for several degenerative conditions due to their multipotency, immunoprivileged properties, and easy expansion in vitro. However, the limited life span of primary CESCs during in vitro expansion greatly hampers their use in basic research and clinical applications. Immortalization of CESCs will overcome this problem and may provide a very useful tool with which to study CESCs biology. Materials and Methods Cells isolated from CEP were cultured in a three-dimensional agarose suspension to acquire proliferative cell clusters. As previous method, we acquire primary CESCs. Then with these cells we showed that silencing p53 expression with lentivirus-mediated small interfering RNA and overexpressing human telomerase reverse transcriptase (hTERT). Results The effects of p53 knockdown and hTERT overexpression on CESCs, including proliferation, colony formation, and differentiation, were determined. The resultant immortal CESCs displayed similar surface antigen profile to primary CESCs and retained CESCs differentiation potential. Gene expression profile showed high similarity between immortalized CESCs and primary CESCs. In addition, immortalization-associated genes were also identified. Conclusion This study provides a new stem cell resources for basic studies of characteristics of CESCs and understanding the mechanism of disc degeneration. Disclosure of Interest None declared