Regulation of differentiation and proliferation of epiphysis stem cells by Notch1 signaling system

• Published in: Zhong hua yi xue za zhi Vol. 85; no. 48; p. 3430
• Main Authors: Zhang, Yi-bei, Chen, An-min, Guo, Feng-jing, Huang, Shi-long, Xiong, Wen-hua, Li, Qi
• Format: Journal Article
• Language: Chinese
• Published: China 21.12.2005
• Subjects: Animals; Cell Differentiation; Cell Proliferation; Cells, Cultured; Collagen Type II - metabolism; Collagen Type X - metabolism; Gene Expression - drug effects; Immunohistochemistry; Peptides - pharmacology; Proliferating Cell Nuclear Antigen - metabolism; Rats; Rats, Sprague-Dawley; Receptor, Notch1 - genetics; Receptor, Notch1 - physiology; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction - drug effects; Stem Cells - cytology; Stem Cells - drug effects
• ISSN: 0376-2491

To investigate the regulation of differentiation and proliferation of epiphysis stem cells by Notch1 signaling system. Costocostal cartilage was taken from a SD rat. Epiphysis stem cells were isolated and cultured. Recombinant human nuclear factor-kappaB (rhNF-kappaB), an activator of the Notch signaling system, and gamma-secretase inhibitor (MW167), an inhibitor of the Notch signaling system, were added into the culture medium respectively. The cells cultured in the medium added with phosphate-buffered saline were used as control group. Then the cultured cells were collected. The expression of the homologous Notch receptors and homologous Notch ligands was detected by RT-PCR. Immunohistochemistry was used to detect the levels of collagen II, collagen X, and proliferating cell nuclear antigen (PCNA). MTT method was used to calculate the growth curve. The cell phase was examined by flow cytometry. The level of alkaline phosphatase (AP) was measured. Western blotting was used to detect the protein expression of