Construction of in vitro models of human bone marrow-derived mesenchymal stem cells differentiated into nucleus pulposus-like cells in rabbits
10.3969/j.issn.1673-8225.2009.45.030
- VernacularTitle:构建兔髓核细胞诱导人骨髓间充质干细胞向类髓核细胞分化的体外模型
- Author:
Quanxiu LI
;
Bohua CHEN
;
Yong LIU
;
Ling XU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(45):8961-8965
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:There is notan effective method for inducing bone marrow-derived mesenchymal stem cells (BMSCs) differentiatied in to intervertebral disc cells.OBJECTIVE:To establish a model of human BMSCs (hBMSCs) differentiated in to nucleus pulposus-like cells in vitro.DESIGN,TIME AND SETTING:A single sample contrast.The experiment was performed at the Orthopaedics Institute and Central Laboratory,the Affiliated Hospital of Qingdao University Medical College,from August 2008 to March 2009.MATERIALS:Two New Zealand white rabbits with 8-week-old were used to culture nucleus pulposus cells.Human nucleus pulposus was obtained from a aged 16 years patient with T_(12)L_1,L_(1.2) congenital scoliosis.The informed consent was obtained from his patients,hBMSCs were purchased from Salye Biological Science and Technology Co.,Ltd.(Guangzhou,China).METHODS:hBMSCs and nucleus pulposus cells of rabbit were co-cultured in new-style culture model (in 6-well plates and inserts with track-etched membrane having 0.4 μm pores without arms) for 7 days,conventionally laminar culture,solitary hBMSCs and nucleus pulpesus cells culture were set up as controls at same time.MAIN OUTCOME MEASURES:The expressions of collagen Ⅱ and aggrecan mRNA were detected by RT-PCR and Western blot.RESULTS:hBMSCs in the conventionally laminar and single hBMSCs groups were negative to collagen Ⅱ and aggrecan.The collagen Ⅱ and aggrecan expression of hBMSCs were upregulated evidently in new-style culture group,which was similar to nucleus pulposus cells that from degenerate intervertebral disc of human,but these expression did not appear in conventionally laminar culture and solitary hBMSCs culture (P ＜ 0.01).The difference among new-style culture group and other groups had no significance (P ＞ 0.05).CONCLUSION:A new-style model of hBMSCs differentiation to nucleus pulposus-like cells is constructed in vitro,which can highly express collagen Ⅱ and aggrecan and provide a foundation for intervertebral disc degeneration cellular treatment.
