Rabbit nucleus pulposus cells induce the differentiation of mesenchymal stem cells after co-culture in a three-dimensional environment
10.3969/j.issn.1673-8225.2009.49.039
- VernacularTitle:三维共培养免髓核细胞诱导骨髓间充质干细胞分化
- Author:
Wenjian CHEN
;
Feng LI
;
Jing YANG
;
Song ZHOU
;
Zhong FANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(49):9783-9786
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: In the field of intervertebral disc tissue engineering, seed cells primarily consist of nucleus pulposus cells (NPCs) and mesenchymal stem cells (MSCs). NPCs have been known to have several shortcomings: limited source, inconvenient collection, poor proliferative capacity, and difficult in vitro culture.OBJECTIVE: To investigate the proliferation of MSCs after co-culture with NPCs in alginate beads-simulated three-dimensional environment.DESIGN, TIME AND SETTING: A cell observation was performed at the Laboratory of Orthopedics, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology between December 2007 and October 2008.MATERIALS: Six healthy New Zealand rabbits of 4 months old were provided by the Laboratory Animal Center, Tongji Medical College of Huazhong University of Science and Technology and were used for the present study.METHODS: Rabbit MSCs were isolated and purified using density gradient centrifugation and adherence methods. Rabbit NPCs were isolated and purified using collagenase Ⅱ digestion and adherence methods. Following liposome-mediated green fluorescent protein tranfection and G418 screening, MSCs of passage 3 were cultured either alone (control group) or with NPCs at a ratio of 1:1 (co-culture group) in alginate beads. After 14 days of culture, alginate beads were dissolved and MSCs were collected by flow cytometry sorting.MAIN OUTCOME MEASURES: The expression of collagen Ⅱ and aggrecan in MSCs was detected by RT-PCR and Western blot techniques.RESULTS: mRNA and protein expression of collagen Ⅱ and aggrecan was observed in the co-culture group, but not in the control group, after 14 days of culture.CONCLUSION: MSC differentiation towards nucleus pulposus-like cells can be induced by co-culture with NPCs in a three-dimensional environment.
