脊索細胞培養基促進BMSCs 增殖分化的研究_《中國修復重建外科雜志》

作者：

馬凱歌 , 邵增務 , 王佰川 , 張彥男 , 楊述華 , 劉濤

華中科技大學同濟醫學院附屬協和醫院骨科（武漢，430022）;

關鍵詞：

組織工程 BMSCs 脊索細胞椎間盤退變兔

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目的為研究椎間盤組織工程種子細胞，觀察脊索細胞培養基對BMSCs 增殖分化的影響。
方法取4 周齡日本大耳白兔胸腰段椎間盤分離培養脊索細胞，取雙側股骨分離培養BMSCs，用含15%FBS 的DMEM/
F12 培養基培養脊索細胞，5 d 后制備脊索細胞培養基。實驗分為兩組，實驗組BMSCs 中加入脊索細胞培養基培養，對照
組BMSCs 中加入含15%FBS 的DMEM/F12 培養基培養。使用細胞活力細胞毒性檢測檢測細胞增殖情況，采用免疫熒光
及實時熒光定量PCR 檢測BMSCs 蛋白多糖及Ⅱ型膠原表達情況。結果成功分離脊索細胞及BMSCs。細胞增殖檢
測示，培養5、7、9、14 d，實驗組細胞數量明顯多于對照組（P lt; 0.05）。免疫熒光檢測示對照組培養7、14 d 細胞內均無或
者有較少Ⅱ型膠原及蛋白多糖表達，實驗組二者均有較多表達，且培養14 d 時表達明顯多于7 d。實時熒光定量PCR 檢
測示，培養7、14 d 實驗組蛋白多糖和Ⅱ型膠原mRNA 表達均顯著高于對照組（P lt; 0.05）；實驗組14 d 蛋白多糖和Ⅱ型膠
原mRNA 表達顯著高于7 d（P lt; 0.05）。結論脊索細胞培養基可促進BMSCs 增殖，并誘導BMSCs 向類軟骨細胞分化，
為脊索細胞和BMSCs 作為種子細胞治療椎間盤退變提供了依據。

引用本文： 馬凱歌,邵增務,王佰川,張彥男,楊述華,劉濤. 脊索細胞培養基促進BMSCs 增殖分化的研究. 中國修復重建外科雜志, 2012, 26(5): 601-606. doi: 復制

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12.	嚴慧深, 吳小濤, 王運濤, 等. 骨髓間充質干細胞修復退變椎間盤中營養和分化效應的變化. 中華實驗外科雜志, 2009, 26(6): 704-706.
13.	Blanco JF, Graciani IF, Sanchez-Guijo FM, et al. Isolation and characterization of mesenchymal stromal cells from human degenerated nucleus pulposus. Spine (Phila Pa 1976), 2010, 35(26): 2259-2265.
14.	Chen J, Yan W, Setton LA. Molecular phenotypes of notochordal cells purified from immature nucleus pulposus. Eur Spine J, 2006, 15 Suppl 3: S303-311.
15.	趙獻峰, 劉浩, 豐干均, 等. 脊索細胞促進髓核軟骨樣細胞增殖及表型維持. 中國修復重建外科雜志, 2008, 22(8): 939-943.
16.	Yang Z, Schmitt JF, Lee EH. Immunohistochemical analysis of human mesenchymal stem cells differentiating into chondrogenic, osteogenic, and adipogenic lineages. Methods Mol Biol, 2011, 698(1): 353-366.
17.	Yang SH, Wu CC, Shih TT, et al. In vitro study on interaction between human nucleus pulposus cells and mesenchymal stem cells through paracrine stimulation. Spine (Phila Pa 1976), 2008, 33(18): 1951-1957.
18.	Strassburg S, Richardson SM, Freemont AJ, et al. Co-culture induces mesenchymal stem cell differentiation and modulation of the degenerate human nucleus pulposus cell phenotype. Regen Med, 2010, 5(5): 701-711.
19.	Erwin WM, Inman RD. Notochord cells regulate intervertebral disc chondrocyte proteoglycan production and cell proliferation. Spine (Phila Pa 1976), 2006, 31(10): 1094-1099.
20.	Steck E, Bertram H, Abel R, et al. Induction of intervertebraldisc-like cells from adult mesenchymal stem cells. Stem Cells, 2005, 23(3): 403-411.
21.	Ivkovic S, Yoon BS, Popoff SN, et al. Connective tissue growth factor coordinates chondrogenesis and angiogenesis during skeletal development. Development, 2003, 130(12): 2779-2791.
22.	Korecki CL, Taboas JM, Tuan RS, et al. Notochordal cell conditioned medium stimulates mesenchymal stem cell differentiation toward a young nucleus pulposus phenotype. Stem Cell Res Ther, 2010, 1(2): 18.
23.	Purmessur D, Schek RM, Abbott RD, et al. Notochordal conditioned media from tissue increases proteoglycan accumulation and promotes a healthy nucleus pulposus phenotype in human mesenchymal stem cells. Arthritis Res Ther, 2011, 13(3): R81.

1. Petit A, Yao G, Rowas SA, et al. Effect of synthetic link N peptide on the expression of type I and type II collagens in human intervertebral disc cells. Tissue Eng Part A, 2011, 17(7-8): 899-904.
2. Hohaus C, Ganey TM, Minkus Y, et al. Cell transplantation in lumbar spine disc degeneration disease. Eur Spine J, 2008, 17(Suppl 4): S492-503.
3. Bertolo A, Thiede T, Aebli N, et al. Human mesenchymal stem cell co-culture modulates the immunological properties of human intervertebral disc tissue fragments in vitro. Eur Spine J, 2011, 20(4): 592-603.
4. Omlor GW, Nerlich AG, Wilke HJ, et al. A new porcine in vivo animal model of disc degeneration: response of anulus fibrosus cells, chondrocyte-like nucleus pulposus cells, and notochordal nucleus pulposus cells to partial nucleotomy. Spine (Phila Pa 1976), 2009, 34(25): 2730-2739.
5. Paesold G, Nerlich AG, Boos N. Biological treatment strategies for disc degeneration: potentials and shortcomings. Eur Spine J, 2007, 16(4): 447-468.
6. Baksh D, Song L, Tuan RS. Adult mesenchymal stem cells: characterization, differentiation, and application in cell and gene therapy. J Cell Mol Med, 2004, 8(3): 301-316.
7. Aguiar DJ, Johnson SL, Oegema TR. Notochordal cells interact with nucleus pulposus cells: regulation of proteoglycan synthesis. Exp Cell Res, 1999, 246(1): 129-137.
8. Johnson WE, Evans H, Menage J, et al. Immunohistochemical detection of Schwann cells in innervated and vascularized human intervertebral discs. Spine (Phila Pa 1976), 2001, 26(23): 2550-2557.
9. Erwin WM, Las Heras F, Islam D, et al. The regenerative capacity of the notochordal cell: tissue constructs generated in vitro under hypoxic conditions. J Neurosurg Spine, 2009, 10(6): 513-521.
10. Risbud MV, Schaer TP, Shapiro IM. Toward an understanding of the role of notochordal cells in the adult intervertebral disc: from discord to accord. Dev Dyn, 2010, 239(8): 2141-2148.
11. Chai C, Leong KW. Biomaterials approach to expand and direct differentiation of stem cells. Mol Ther, 2007, 15(3): 467-480.
12. 嚴慧深, 吳小濤, 王運濤, 等. 骨髓間充質干細胞修復退變椎間盤中營養和分化效應的變化. 中華實驗外科雜志, 2009, 26(6): 704-706.
13. Blanco JF, Graciani IF, Sanchez-Guijo FM, et al. Isolation and characterization of mesenchymal stromal cells from human degenerated nucleus pulposus. Spine (Phila Pa 1976), 2010, 35(26): 2259-2265.
14. Chen J, Yan W, Setton LA. Molecular phenotypes of notochordal cells purified from immature nucleus pulposus. Eur Spine J, 2006, 15 Suppl 3: S303-311.
15. 趙獻峰, 劉浩, 豐干均, 等. 脊索細胞促進髓核軟骨樣細胞增殖及表型維持. 中國修復重建外科雜志, 2008, 22(8): 939-943.
16. Yang Z, Schmitt JF, Lee EH. Immunohistochemical analysis of human mesenchymal stem cells differentiating into chondrogenic, osteogenic, and adipogenic lineages. Methods Mol Biol, 2011, 698(1): 353-366.
17. Yang SH, Wu CC, Shih TT, et al. In vitro study on interaction between human nucleus pulposus cells and mesenchymal stem cells through paracrine stimulation. Spine (Phila Pa 1976), 2008, 33(18): 1951-1957.
18. Strassburg S, Richardson SM, Freemont AJ, et al. Co-culture induces mesenchymal stem cell differentiation and modulation of the degenerate human nucleus pulposus cell phenotype. Regen Med, 2010, 5(5): 701-711.
19. Erwin WM, Inman RD. Notochord cells regulate intervertebral disc chondrocyte proteoglycan production and cell proliferation. Spine (Phila Pa 1976), 2006, 31(10): 1094-1099.
20. Steck E, Bertram H, Abel R, et al. Induction of intervertebraldisc-like cells from adult mesenchymal stem cells. Stem Cells, 2005, 23(3): 403-411.
21. Ivkovic S, Yoon BS, Popoff SN, et al. Connective tissue growth factor coordinates chondrogenesis and angiogenesis during skeletal development. Development, 2003, 130(12): 2779-2791.
22. Korecki CL, Taboas JM, Tuan RS, et al. Notochordal cell conditioned medium stimulates mesenchymal stem cell differentiation toward a young nucleus pulposus phenotype. Stem Cell Res Ther, 2010, 1(2): 18.
23. Purmessur D, Schek RM, Abbott RD, et al. Notochordal conditioned media from tissue increases proteoglycan accumulation and promotes a healthy nucleus pulposus phenotype in human mesenchymal stem cells. Arthritis Res Ther, 2011, 13(3): R81.

《中國修復重建外科雜志》

脊索細胞培養基促進BMSCs 增殖分化的研究

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《中國修復重建外科雜志》

脊索細胞培養基促進BMSCs 增殖分化的研究

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

Format

Content

摘要全文圖表視頻參考文獻施引文獻補充材料