Induced differentiation of nucleus pulposus cells from bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2013.06.008

Abstract

Abstract:

BACKGROUND: There have been many reports addressing repair of degenerative lumbar intervertebral discs in rats using alginate compounded with bone marrow mesenchymal stem cells. However, there are few reports describing biological indices of this compounded material versus normal nucleus pulposus cells.
OBJECTIVE: To investigate the inducing effects of normal nucleus pulposus cells on bone marrow mesenchymal stem cells under Transwell non-contact co-culture condition.
METHODS: Rat bone marrow mesenchymal stem cells were cultured from the whole bone marrow and identified. Passage 3 bone marrow mesenchymal stem cells and nucleus pulposus cells compounded onto alginate were inoculated into a Transwell co-culture system. Bone marrow mesenchymal stem cells compounded onto alginate were inoculated onto the upper layer, and nucleus pulposus cells compounded onto alginate were inoculated onto the lower layer. After 7 days of co-culture, bone marrow mesenchymal stem cells were retrieved from the upper layer.
RESULTS AND CONCLUSION: RT-PCR results showed that type Ⅱ collagen, Sox-9 and versian mRNA expression in the bone marrow mesenchymal stem cells + nucleus pulposus cells group was similar to that in the normal nucleus pulposus cells group. These findings suggest that after co-cultured with nucleus pulposus cells, bone marrow mesenchymal stem cells can be induced to differentiate into nucleus pulposus cells.

Key words: stem cells, bone marrow-derived stem cells, sox9, bone marrow mesenchymal stem cells, alginate, nucleus pulposus, RT-PCR, intervertebral disc degeneration, type Ⅱ collagen, version, Transwell co-culture system, other grants-supported paper, stem cell photographs-containing paper

CLC Number:

R394.2

Chen Liang, Li Da-peng, Huang Yong-hui, Liu Kan. Induced differentiation of nucleus pulposus cells from bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(6): 992-998.

Figures/Tables 11

References

[1] Vadalà G, Studer RK, Sowa G, et al. Coculture of bone marrow mesenchymal stem cells and nucleus pulposus cells modulate gene expression profile without cell fusion.Spine (Phila Pa 1976). 2008;33(8):870-876.

[2] Diwan AD, Parvataneni HK, Khan SN,et al. Current concepts in intervertebral disc restoration. Orthop Clin North Am. 2000; 31(3):453-464.

[3] Liu ZJ, Zhuge Y, Velazquez OC.Trafficking and differentiation of mesenchymal stem cells. J Cell Biochem. 2009;106(6): 984-991.

[4] Xu CF,Hu DH,Zhao ZT,et al. Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2010; 14(6):1002-1005. 徐成峰,胡大海,赵周婷,等.兔骨髓间充质干细胞体外分离培养及多向诱导分化[J].中国组织工程研究与临床康复,2010, 14(6): 1002-1005.

[5] Gun QY,Lu SB,Zhang L.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2006;10(1):61-63. 郭全义,卢世璧,张莉.藻酸钙凝胶为载体体外短期培养羊软骨细胞的生物学性状[J].中国组织工程研究与临床康复,2006,10(1): 61-63.

[6] Gomes ME, Bossano CM, Johnston CM, et al. In vitro localization of bone growth factors in constructs of biodegradable scaffolds seeded with marrow stromal cells and cultured in a flow perfusion bioreactor.Tissue Eng. 2006; 12(1):177-188.

[7] Cui L, Wu Y, Cen L, et al. Repair of articular cartilage defect in non-weight bearing areas using adipose derived stem cells loaded polyglycolic acid mesh. Biomaterials. 2009;30(14): 2683-2693.

[8] Zhu XY, Zhang XZ, Xu L, et al. Transplantation of adipose- derived stem cells overexpressing hHGF into cardiac tissue. Biochem Biophys Res Commun. 2009;379(4):1084-1090.

[9] Sakai D, Mochida J, Yamamoto Y,et al. Transplantation of mesenchymal stem cells embedded in Atelocollagen gel to the intervertebral disc: a potential therapeutic model for disc degeneration. Biomaterials. 2003;24(20):3531-3541.

[10] Chen WJ,Li F,Yang J,et al. Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2009;13(49):9783-9786. 陈文坚,李锋,杨晶,等.三维共培养兔髓核细胞诱导骨髓间充质干细胞分化[J].中国组织工程研究与临床康复,2009,13(49): 9783-9786.

[11] Tian HQ,Li F. Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2009;13(38):7417-7421. 田海泉,李放.体外三维支架诱导脂肪干细胞向髓核样细胞的分化[J].中国组织工程研究与临床康复,2009,13(38):7417-7421.

[12] Perka C, Spitzer RS, Lindenhayn K,et al. Matrix-mixed culture: new methodology for chondrocyte culture and preparation of cartilage transplants. J Biomed Mater Res. 2000;49(3): 305-311.

[13] Ueng SW, Yuan LJ, Lee N,et al. In vivo study of biodegradable alginate antibiotic beads in rabbits. J Orthop Res. 2004;22(3):592-599.

[14] Steck E, Fischer J, Lorenz H, et al. Mesenchymal stem cell differentiation in an experimental cartilage defect: restriction of hypertrophy to bone-close neocartilage. Stem Cells Dev. 2009;18(7):969-978.

[15] Getgood A, Brooks R, Fortier L, et al. Articular cartilage tissue engineering: today's research, tomorrow's practice. J Bone Joint Surg Br. 2009;91(5):565-576.

[16] Mithoefer K, McAdams TR, Scopp JM, et al. Emerging options for treatment of articular cartilage injury in the athlete. Clin Sports Med. 2009;28(1):25-40.

[17] Markusen JF, Mason C, Hull DA, et al. Behavior of adult human mesenchymal stem cells entrapped in alginate-GRGDY beads. Tissue Eng. 2006;12(4):821-830.

[18] Wang WZ,Chen ZX,Liu XQ. Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2010;14(38):7041-7044. 王万宗,陈宗雄,刘晓强.骨髓间充质干细胞与关节软骨细胞Transwell共培养诱导形成软骨[J].中国组织工程研究与临床康复,2010,14(38):7041-7044.

[19] Bi W, Deng JM, Zhang Z, et al. Sox9 is required for cartilage formation. Nat Genet. 1999;22(1):85-89.