Differentiation Of Mouse Parthenogenetic Embryonic Stem Cells Into The Chondrogenic Lineage In Vitro

Embryonic stem cells (ESCs) have the potential to offer an unlimited source of chondrogenic cells for application in cartilage repair and regeneration. However, ethical controversy still ramains as an obstacle over their application. Recent studies have demonstrated the multiple differentiation potentials of parthenogenetic embryonic stem cells (pESCs) derived from parthenogenetic embryos, which have relatively low immune rejection and will die after approximately 10 d of gestation, due to the absence of paternal genes and the limited development of extra-embryonic tissues. pESCs could develop into components of all three germ layers, providing the possibility to induce them to differentiate into chondrogenic lineage.In this study, we hypothesized that mouse pESCs could act as seed cells for cartilage tissue engineering. After the investigation of pluripotency, ability of proliferation, gene expression patterns, formation of embryoid bodies (EB) in vitro and teratomas in vivo, we employed the strategy of high-density micromass model to direct mouse pESCs to differentiate into chondrogenic lineage under the presence of BMP-2 and TGF-β.And then cultured the prechondrogenic cells as pellet for a more adequately differentiation. Chondrogenic differentiation were demonstrated by real-time PCR measurement of cartilage-related gene expression, histological and immunofluorescent staining of matrix protein synthesis. The results showed that the expressions of cartilage-specific gene were significantly upregulated, with the synthesis secretion and deposition of matrix protein including proteoglycan and Col Ⅱ.21 days later, we obtained cartilage like tissue samples with glistering white, smooth surface and certain mechanical strength.Here we demonstrated that mouse pESC could differentiate into chondrogenic cells, and may provide a practical and ethical-unlimited strategy for cartilage tissue engineering.