The effects of CD44 fucosylation on fluid adhesion force of rabbit bone marrow mesenchymal stem cells_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

XIAO Fei ,  JIAO Jing , HUANG Yucheng , XU Haijun , ZUO Wei , WANG Junwen

Department of Orthopedics, Wuhan Puai Hospital, Wuhan Hubei, 430000, P.R.China;

Corresponding?author：

JIAO Jing, Email: realzebra@qq.com

Keywords：

Bone marrow mesenchymal stem cells; fucosylation; E-selectin; fluid adhesion force; rabbit

DOI：

10.7507/1002-1892.201704012

Video：

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Objective To investigate the effect of CD44 fucosylation on fluid adhesion force of rabbit bone marrow mesenchymal stem cells (BMSCs). Methods The rabbit BMSCs were isolated and purified by density gradient centrifugation combined with adherent culture method. The morphology of cells were observed by inverted microscope, and the cell surface markers of CD44, CD34, CD29, and CD105 were assessed by flow cytometry. BMSCs fucosylated by alpha-(1, 3)-fucosyltransferase Ⅵ (FTⅥ) were as the experimental group, and the non-fucosylated BMSCs were as the control group, and then the positive rate of sialyl-LewisX (sLe^X) and the binding rate of E-selectin were detected by flow cytometry. The fucosylated BMSCs resuspended in Hank balanced salt solution (HBSS) were assigned as the experimental group (group A), at same time, the non-fucosylated BMSCs resuspended in HBSS solution as the study control group (group B), and the fucosylated BMSCs resuspended in HBSS solution which was added EDTA as negative control group (group C). The fluid adhesion force of rabbit BMSCs were detected by the parallel flow chamber adhesion test. Results Primary BMSCs mainly shaped as spindle and kept strong growth. The third generation BMSCs were negative for CD34, but positive for CD44, CD29, and CD105. After fucosylation, the positive rate of sLe^X in the experimental group was 32.52%±1.76%, which was significantly higher than that in the control group (1.48%±0.51%) (t=29.277, P= 0.000). The binding rate of E-selectin in the experimental group was 41.05%±1.84%, which was also significantly higher than that in the control group (4.33%±0.92%) (t=35.674, P=0.000). With the increase of fluid shear force, the number of BMSCs adhering to the surface of human umbilical vascular endothelial cells (HUVEC) in group A was increased at first and then decreased, while there was few BMSCs adhering to the surface of HUVEC in groups B and C. Under the different fluid shear stress, the number of BMSCs adhered to the surface of HUVEC in group A was significantly higher than that in groups B and C (P<0.05), and there was no significant difference between groups B and C (P>0.05). Conclusion CD44 fucosylation on BMSCs can enhance the fluid adhesion force of rabbit BMSCs.

Citation： XIAO Fei, JIAO Jing, HUANG Yucheng, XU Haijun, ZUO Wei, WANG Junwen. The effects of CD44 fucosylation on fluid adhesion force of rabbit bone marrow mesenchymal stem cells. Chinese Journal of Reparative and Reconstructive Surgery, 2018, 32(1): 99-103. doi: 10.7507/1002-1892.201704012 Copy

1.	徐紅珍, 孫俊, 蘇儉生. 兔骨髓間充質干細胞的分離、培養與鑒定. 同濟大學學報 (醫學版) , 2010, 31(5): 16-20.
2.	榮為為, 韓明子, 金世柱, 等. 骨髓間充質干細胞在組織工程研究中的應用新進展. 現代生物醫學進展, 2017, 17(5): 982-984.
3.	Weinand C, Neville CM, Weinberg E, et al. Optimizing biomaterials for tissue engineering human bone using mesenchymal stem cells. Plast Reconstr Surg, 2016, 137(3): 854-863.
4.	劉陽, 朱立新, 楊宏, 等. 可注射性納米羥基磷灰石/殼聚糖復合骨髓間充質干細胞促進骨缺損的修復. 中國組織工程研究, 2010, 14(34): 6278-6282.
5.	程中華, 薛威, 王李琴, 等. 骨髓間充質干細胞結合硼硅酸鹽玻璃支架修復兔股骨頭壞死. 骨科, 2017, 8(2): 121-126.
6.	Mortezaee K, Pasbakhsh P, Kashani IR, et al. Melatonin pretreatment enhances the homing of bone marrow-derived mesenchymal stem cells following transplantation in a rat model of liver fibrosis. Iran Biomed J, 2016, 20(4): 207-216.
7.	Si X, Liu X, Li J, et al. Transforming growth factor-β1 promotes homing of bone marrow mesenchymal stem cells in renal ischemia-reperfusion injury. Int J Clin Exp Pathol, 2015, 8(10): 12368-12378.
8.	Zimolag E, Borowczyk-MJ, Kedracka-KS, et al. Electric field as a potential directional cue in homing of bone marrow-derived mesenchymal stem cells to cutaneous wounds. Biochim Biophys Acta, 2017, 1864(2): 267-279.
9.	Sackstein R. Glycoengineering of HCELL, the human bone marrow homing receptor: sweetly programming cell migration. Ann Biomed Eng, 2012, 40(4): 766-776.
10.	Sackstein R, Merzaban JS, Cain DW, et al. Ex vivo glycan engineering of CD44 programs human multipotent mesenchymal stromal cell trafficking to bone. Nat Med, 2008, 14(2): 181-187.
11.	Chou KJ, Lee PT, Chen CL, et al. CD44 Fucosylation on mesenchymal stem cell enhances homing and macrophage polarization in ischemic kidney injury. Exp Cell Res, 2017, 350(1): 91-102.
12.	Dykstra B, Lee J, Mortensen LJ, et al. Glycoengineering of e-selectin ligands by intracellular versus extracellular fucosylation differentially affects osteotropism of human mesenchymal stem cells. Stem Cells, 2016, 34(10): 2501-2511.
13.	Dimitroff CJ, Lee JY, Rafii S, et al. CD44 Is a major e-selectin ligand on human hematopoietic progenitor cells. J Cell Biol, 2001, 153(6): 1277-1286.
14.	Krause DS, Lazarides K, Andrian UH, et al. Requirement for CD44 in homing and engraftment of BCR-ABL-expressing leukemic stem cells. Nat Med, 2006, 12(10): 1175-1180.
15.	Antoine M, Tag CG, Gressner AM, et al. Expression of E-selectin ligand-1 (CFR/ESL-1) on hepatic stellate cells: implications for leukocyte extravasation and liver metastasis. Oncol Rep, 2009, 21(2): 357-362.
16.	Xia L, McDaniel JM, Yago T, et al. Surface fucosylation of human cord blood cells augments binding to P-selectin and E-selectin and enhances engraftment in bone marrow. Blood, 2004, 104(10): 3091-3096.
17.	曹華, 高建華, 劉小龍, 等. 大鼠骨髓間充質干細胞分離鑒定方法及生物學特性. 中國組織工程研究, 2017, 21(9): 1357-1361.
18.	羅世君, 吳倩倩, 孫勇, 等. 兔骨髓間充質干細胞的分離培養及成骨分化的實驗研究. 西南國防醫藥, 2016, 26(6): 590-593.
19.	Pachón-Pe?a G, Donnelly C, Ruiz-Ca?ada C, et al. A glycovariant of human CD44 is characteristically expressed on human mesenchymal stem cells. Stem Cells, 2017, 35(4): 1080-1092.
20.	Chou KJ, Lee PT, Chen CL, et al. CD44 fucosylation on mesenchymal stem cell enhances homing and macrophage polarization in ischemic kidney injury. Exp Cell Res, 2017, 350(1): 91-102.

1. 徐紅珍, 孫俊, 蘇儉生. 兔骨髓間充質干細胞的分離、培養與鑒定. 同濟大學學報 (醫學版) , 2010, 31(5): 16-20.
2. 榮為為, 韓明子, 金世柱, 等. 骨髓間充質干細胞在組織工程研究中的應用新進展. 現代生物醫學進展, 2017, 17(5): 982-984.
3. Weinand C, Neville CM, Weinberg E, et al. Optimizing biomaterials for tissue engineering human bone using mesenchymal stem cells. Plast Reconstr Surg, 2016, 137(3): 854-863.
4. 劉陽, 朱立新, 楊宏, 等. 可注射性納米羥基磷灰石/殼聚糖復合骨髓間充質干細胞促進骨缺損的修復. 中國組織工程研究, 2010, 14(34): 6278-6282.
5. 程中華, 薛威, 王李琴, 等. 骨髓間充質干細胞結合硼硅酸鹽玻璃支架修復兔股骨頭壞死. 骨科, 2017, 8(2): 121-126.
6. Mortezaee K, Pasbakhsh P, Kashani IR, et al. Melatonin pretreatment enhances the homing of bone marrow-derived mesenchymal stem cells following transplantation in a rat model of liver fibrosis. Iran Biomed J, 2016, 20(4): 207-216.
7. Si X, Liu X, Li J, et al. Transforming growth factor-β1 promotes homing of bone marrow mesenchymal stem cells in renal ischemia-reperfusion injury. Int J Clin Exp Pathol, 2015, 8(10): 12368-12378.
8. Zimolag E, Borowczyk-MJ, Kedracka-KS, et al. Electric field as a potential directional cue in homing of bone marrow-derived mesenchymal stem cells to cutaneous wounds. Biochim Biophys Acta, 2017, 1864(2): 267-279.
9. Sackstein R. Glycoengineering of HCELL, the human bone marrow homing receptor: sweetly programming cell migration. Ann Biomed Eng, 2012, 40(4): 766-776.
10. Sackstein R, Merzaban JS, Cain DW, et al. Ex vivo glycan engineering of CD44 programs human multipotent mesenchymal stromal cell trafficking to bone. Nat Med, 2008, 14(2): 181-187.
11. Chou KJ, Lee PT, Chen CL, et al. CD44 Fucosylation on mesenchymal stem cell enhances homing and macrophage polarization in ischemic kidney injury. Exp Cell Res, 2017, 350(1): 91-102.
12. Dykstra B, Lee J, Mortensen LJ, et al. Glycoengineering of e-selectin ligands by intracellular versus extracellular fucosylation differentially affects osteotropism of human mesenchymal stem cells. Stem Cells, 2016, 34(10): 2501-2511.
13. Dimitroff CJ, Lee JY, Rafii S, et al. CD44 Is a major e-selectin ligand on human hematopoietic progenitor cells. J Cell Biol, 2001, 153(6): 1277-1286.
14. Krause DS, Lazarides K, Andrian UH, et al. Requirement for CD44 in homing and engraftment of BCR-ABL-expressing leukemic stem cells. Nat Med, 2006, 12(10): 1175-1180.
15. Antoine M, Tag CG, Gressner AM, et al. Expression of E-selectin ligand-1 (CFR/ESL-1) on hepatic stellate cells: implications for leukocyte extravasation and liver metastasis. Oncol Rep, 2009, 21(2): 357-362.
16. Xia L, McDaniel JM, Yago T, et al. Surface fucosylation of human cord blood cells augments binding to P-selectin and E-selectin and enhances engraftment in bone marrow. Blood, 2004, 104(10): 3091-3096.
17. 曹華, 高建華, 劉小龍, 等. 大鼠骨髓間充質干細胞分離鑒定方法及生物學特性. 中國組織工程研究, 2017, 21(9): 1357-1361.
18. 羅世君, 吳倩倩, 孫勇, 等. 兔骨髓間充質干細胞的分離培養及成骨分化的實驗研究. 西南國防醫藥, 2016, 26(6): 590-593.
19. Pachón-Pe?a G, Donnelly C, Ruiz-Ca?ada C, et al. A glycovariant of human CD44 is characteristically expressed on human mesenchymal stem cells. Stem Cells, 2017, 35(4): 1080-1092.
20. Chou KJ, Lee PT, Chen CL, et al. CD44 fucosylation on mesenchymal stem cell enhances homing and macrophage polarization in ischemic kidney injury. Exp Cell Res, 2017, 350(1): 91-102.

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