[Promotion effect of notochordal cells conditioned medium on proliferation and differentiation of bone marrow mesenchymal stem cells].
OBJECTIVE
To investigate the effect of notochordal cells (NCs) conditioned medium (NCCM) on the proliferation and differentiation of bone marrow mesenchymal stem cells (BMSCs).


METHODS
NCs and BMSCs were isolated from the thoracolumbar intervertebral disc and the femurs of 4-week-old Japanese white rabbits, respectively. NCs were cultured with DMEM/F12 medium containing 15% FBS for 5 days to prepare NCCM. The experiment consisted of 2 groups: BMSCs were cultured with NCCM in experimental group and with DMEM/F12 medium containing 15% FBS in control group. The proliferation of BMSCs was assessed by cell counting kit-8 at 1, 3, 5, 7, 9, and 14 days after culture; the expression of proteoglycan and collagen type II were determined by immunofluorescence and real-time fluorescent quantitative PCR at 7 and 14 days after culture.


RESULTS
NCs and BMSCs were successfully isloated. At 5, 7, 9, and 14 days, the number of BMSCs in the experimental group was significantly more than those in the control group (P < 0.05). At 7 and 14 days, there was no expression or less expression of proteoglycan and collagen type II in the control group; however, there was a lot of expression of proteoglycan and collagen type II in the experimental group, and the expressions were higher at 14 days than at 7 days. At 7 and 14 days after culture, the mRNA expressions of proteoglycan and collagen type II were significantly higher in the experimental group than in the control group (P < 0.05), and at 14 days than at 7 days in the experimental group (P < 0.05).


CONCLUSIONS
NCCM can promote the proliferation and the differentiation of BMSCs into chondroyte-like cells, which provides the basis for NCs and BMSCs as seed cells in the treatment of degenerative disc disease.
Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan Hubei 430022, PR China.
Kaige
K
Ma
Zengwu
Z
Shao
Baichuan
B
Wang
Yannan
Y
Zhang
Shuhua
S
Yang
Tao
T
Liu
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000378:metabolism / Q000494:pharmacology*; D005260:Female; D007403:Intervertebral Disc / Q000166:cytology; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D009672:Notochord / Q000166:cytology*; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D023822:Tissue Engineering / Q000379:methods
D024043:Collagen Type II; D017077:Culture Media, Conditioned; D011509:Proteoglycans; D012333:RNA, Messenger