IMMUNOGENICITY OF HUMAN UMBILICAL CORD BLOOD DERIVED MESENCHYMAL STEM CELLS AFTER OSTEOGENIC INDUCTION_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

YANGDawei ¹ , LINHemin ² ,  LIUGuangpeng ³

1. Department of Orthopaedic Surgery, the Fourth Hospital of Harbin Medical University, Harbin Heilongjiang, 150001, P. R. China;
2. Department of Plastic and Reconstructive Surgery, Eye Hospital Affiliated to Wenzhou Medical University;
3. Department of Plastic and Reconstructive Surgery, Shanghai Tenth People's Hospital, Tongji University;

Corresponding?author：

LIUGuangpeng, Email: guangpengliu@163.com

Keywords：

Umbilical cord blood derived mesenchymal stem cells; Osteogenic differentiation; Immunological property; Lymphocyte

DOI：

10.7507/1002-1892.20140167

Video：

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Objective To study the immunological properties of osteogenically differentiated umbilical cord blood derived mesenchymal stem cells (UCB-MSCs). Methods UCB-MSCs were isolated from the umbilical cord vein, and were expanded; the cells at passage 3 were osteogenically induced for 2 weeks in vitro. The expressions of human leukocyte antigen I (HLA-I) and HLA-Ⅱ molecules were observed by flow cytometry analysis before and after osteogenic induction. Peripheral blood T lymphocytes were isolated and cultured with osteoblastic induced or non-osteoblastic induced UCB-MSCs in different cell concentrations of 1×10², 1×10³, 1×10⁴, and 1×10⁵ cells/well. The intake value of 3H-thymidine was calculated with luminescence counter. Then T lymphocytes were pretreated with PHA, and co-cultured with osteoblastic induced and non-osteoblastic induced UCB-MSCs as described above. IL-2 was further added to test the reversed effect of T lymphocytes proliferation stimulated by UCB-MSCs. Finally, to investigate whether the immunomodulatory effects on T lymphocytes proliferation depend on direct or indirect cell contact, the Transwell chamber culture system of UCB-MSCs and T lymphocytes was established. Results Flow cytometry analysis showed that non-osteoblastic induced UCB-MSCs expressed HLA-I but did not express HLA-Ⅱ; the expression of HLA-Ⅱ increased in osteoblastic induced UCB-MSCs. No T lymphocyte response was stimulated by non-osteoblastic induced UCB-MSCs, but osteoblastic induced UCB-MSCs could stimulate the proliferation of allogeneic T lymphocytes, especially after IFN-γ treatment. Non-osteoblastic induced UCB-MSCs of 1×10⁴ and 1×10⁵ cells/well could suppress the proliferation of T lymphocytes evoked by PHA, and this suppression could be reversed by the addition of IL-2. While osteoblastic induced UCB-MSCs did not have such suppressive effect. The results of the Transwell culture system also showed that non-osteoblastic induced UCB-MSCs could obviously inhibit the proliferation of T lymphocytes, but the osteoblastic induced UCB-MSCs could not. Conclusion The immunological properties of UCB-MSCs will change accordingly after osteogenic induction, so UCB-MSCs might not be suitable for the seed cells of bone tissue engineering.

Citation： YANGDawei, LINHemin, LIUGuangpeng. IMMUNOGENICITY OF HUMAN UMBILICAL CORD BLOOD DERIVED MESENCHYMAL STEM CELLS AFTER OSTEOGENIC INDUCTION. Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(6): 752-757. doi: 10.7507/1002-1892.20140167 Copy

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2.	Lee OK, Kuo TK, Chen WM, et al. Isolation of multipotent mesenchymal stem cells from umbilical cord blood. Blood, 2004, 103(5):1669-1675.
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10.	Yoo KH, Jang IK, Lee MW, et al. Comparison of immunomodulatory properties of mesenchymal stem cells derived from adult human tissues. Cell Immunol, 2009, 259(2):150-156.
11.	Wang M, Yang Y, Yang D, et al. The immunomodulatory activity of human umbilical cord bloodderived mesenchymal stem cells in vitro. Immunology, 2008, 126(2):220-232.
12.	Ji F, Wang Y, Sun H, et al. Human umbilical cord blood-derived non-hematopoietic stem cells suppress lymphocyte proliferation and CD4, CD8 expression. J Neuroimmunol, 2008, 197(2):99-109.
13.	Uccelli A, Moretta L, Pistoia V. Immunoregulatory function of mesenchymal stem cells. Eur J Immunol, 2006, 36(10):2566-2673.
14.	李宇琳, 周恒, 劉廣鵬, 等. 茜素紅半定量測定細胞基質鈣含量的實驗研究. 組織工程與重建外科雜志, 2008, 4(2):61-64.
15.	Niemeyer P, Kornacker M, Mehlhorn A, et al. Comparison of immunological properties of bone marrow stromal cells and adipose tissue-derived stem cells before and after osteogenic differentiation in vitro. Tissue Eng, 2007, 13(1):111-121.
16.	Uccelli A, Moretta L, Pistoia V. Mesenchymal stem cells in health and disease. Nat Rev Immunol, 2008, 8(9):726-736.
17.	Niemeyer P, Schonberger TS, Hahn J, et al. Xenogenic transplantation of human mesenchymal stem cells in a critical size defect of the sheep tibia for bone regeneration. Tissue Eng, 2010, 16(1):33-43.
18.	王瓊, 劉婷, 張耀林, 等. 人胎盤MSCs對異基因小鼠皮膚移植排斥的免疫調節作用. 中國修復重建外科雜志, 2013, 27(7):775-780.

1. Yang SE, Ha CW, Jung M, et al. Mesenchymal stem progenitor cells developed in cultures from UC blood. Cytotherapy, 2004, 6(5):476-486.
2. Lee OK, Kuo TK, Chen WM, et al. Isolation of multipotent mesenchymal stem cells from umbilical cord blood. Blood, 2004, 103(5):1669-1675.
3. Kern S, Eichler H, Stoeve J, et al. Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells, 2006, 24(5):1294-1301.
4. Chang YJ, Tseng CP, Hsu LF, et al. Characterization of two populations of mesenchymal progenitor cells in umbilical cord blood. Cell Biol Int, 2006, 30(6):495-499.
5. 劉廣鵬, 張文杰, 李宇琳, 等. 長期體外培養對人臍血間充質干細胞成骨分化潛能的影響. 組織工程與重建外科雜志, 2009, 5(1):4-6.
6. Liu G, Li Y, Sun J, et al. In vitro and in vivo evaluation of osteogenesis of human umbilical cord blood derived mesenchymal stem cells on partially demineralized bone. Tissue Eng Part A, 2010, 16(3):971-982.
7. Campagnoli C, Roberts IA, Kumar S, et al. Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood, 2001, 98(8):2396-2402.
8. Fritsch G, Stimpfl M, Kurz M, et al. The composition of CD34 subpopulations differs between bone marrow, blood and cord blood. Bone Marrow Transplant, 1996, 17(2):169-178.
9. Lu L, Shen RN, Broxmeyer HE. Stem cells from bone marrow, umbilical cord blood and peripheral blood for clinical application:current status and future application. Crit Rev Oncol Hematol, 1996, 22(2):61-78.
10. Yoo KH, Jang IK, Lee MW, et al. Comparison of immunomodulatory properties of mesenchymal stem cells derived from adult human tissues. Cell Immunol, 2009, 259(2):150-156.
11. Wang M, Yang Y, Yang D, et al. The immunomodulatory activity of human umbilical cord bloodderived mesenchymal stem cells in vitro. Immunology, 2008, 126(2):220-232.
12. Ji F, Wang Y, Sun H, et al. Human umbilical cord blood-derived non-hematopoietic stem cells suppress lymphocyte proliferation and CD4, CD8 expression. J Neuroimmunol, 2008, 197(2):99-109.
13. Uccelli A, Moretta L, Pistoia V. Immunoregulatory function of mesenchymal stem cells. Eur J Immunol, 2006, 36(10):2566-2673.
14. 李宇琳, 周恒, 劉廣鵬, 等. 茜素紅半定量測定細胞基質鈣含量的實驗研究. 組織工程與重建外科雜志, 2008, 4(2):61-64.
15. Niemeyer P, Kornacker M, Mehlhorn A, et al. Comparison of immunological properties of bone marrow stromal cells and adipose tissue-derived stem cells before and after osteogenic differentiation in vitro. Tissue Eng, 2007, 13(1):111-121.
16. Uccelli A, Moretta L, Pistoia V. Mesenchymal stem cells in health and disease. Nat Rev Immunol, 2008, 8(9):726-736.
17. Niemeyer P, Schonberger TS, Hahn J, et al. Xenogenic transplantation of human mesenchymal stem cells in a critical size defect of the sheep tibia for bone regeneration. Tissue Eng, 2010, 16(1):33-43.
18. 王瓊, 劉婷, 張耀林, 等. 人胎盤MSCs對異基因小鼠皮膚移植排斥的免疫調節作用. 中國修復重建外科雜志, 2013, 27(7):775-780.

Chinese Journal of Reparative and Reconstructive Surgery

IMMUNOGENICITY OF HUMAN UMBILICAL CORD BLOOD DERIVED MESENCHYMAL STEM CELLS AFTER OSTEOGENIC INDUCTION

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