Immunosuppressive and Hematopoiesis-Supporting Properties of Stromal Cells at Low Oxygen

Elena R Andreeva^1, , Irina V Andrianova¹, Polina I Bobyleva¹ and Ludmila B Buravkova¹

¹Cell Physiology lab, Institute of BioMedical Problems, Russian Academy of Sciences, Moscow, Russia

Cite this paper:
Elena R Andreeva, Irina V Andrianova, Polina I Bobyleva and Ludmila B Buravkova. Immunosuppressive and Hematopoiesis-Supporting Properties of Stromal Cells at Low Oxygen. American Journal of Biomedical Research. 2013; 1(1):7-12. doi: 10.12691/ajbr-1-1-2

Abstract

In adults, low differentiated stromal cells like fibroblasts, multipotent mesenchymal stromal cells, reticular cells etc. are important integrative components of tissue homeostasis and regeneration. The purpose of this study was to compare immunomodulatory activity and hematopoiesis-supportive capacity of two human stromal cell types: fetal fibroblasts (fFBs) and multipotent mesenchymal stromal cells (MMSCs) under different oxygen tension (20% vs. 5%). Surface immunophenotyping by flow cytometry revealed that MMSCs and fFBs were СD90, СD105, HLA-ABC positive and СD34, СD45, HLA-DR negative regardless of the О₂ concentration. The percentage of T-cells was relatively unchanged after 72 hours co-culture with stromal cells; the proportion of B- and NK-cells was increased, but the share of CD3⁺/HLA-DR⁺ cells was decreased when co-culturing with both MMSCs and fFBs. This reduction was statistically significant at 5% O₂ in comparison with 20% O₂ (p < 0.05). CD3⁺/CD25⁺ cells decreased after co-culture with both stromal cells in 20% O₂. This effect was revealed only for MMSC at 5% O₂. After 72 hours in co-culture with MMSCs, mononuclear cells from umbilical cord blood (CBMCs) maintained the same number of colony forming units (CFU) as compared with the initial CBMC suspension. The number of CFU was 30% higher at 5% O₂ than in 20% O₂ (p < 0.05). The CFU number in CBMCs after co-culture with fFBs decreased and a stimulatory effect of low oxygen was not observed. CBMCs that were adhered to MMSCs formed colonies of undifferentiated progenitors after 2 weeks of co-culture both at 20% and 5% O₂. Colonies were not found on fFBs. The immunomodulatory potency and the hematopoiesis-supportive capacity of stromal cells can vary depending on their tissue origin and may be modified by the partial pressure of oxygen.

Keywords:
human multipotent mesenchymal stromal cells from adipose tissue human fetal fibroblasts peripheral blood mononuclear cells mononuclear cells from umbilical cord blood coculture immunosuppression hematopoiesis low oxygen tension

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References:

[1]	Jones, B.J. and McTaggart, S.J., Immunosuppression by mesenchymal stromal cells: from culture to clinic, Exp Hematol, 36(6): 733-741, 2008.
[2]	Buravkova, L.B. and Andreeva, E.R., Interaction of human multipotential mesenchymal stromal and immune cells, Human Physiology, 36(5): 110-120, 2010.
[3]	Chen, P.M., Yen, M.L., Liu, K.J., Sytwum H.K. and Yenm B.L., Immunomodulatory properties of human adult and fetal multipotent mesenchymal stem cells, J Biomed Sci., 18(1): 49, 2011.
[4]	Sarkhosh, K., Tredget, E.E., Karami, A., Uludag, H., Iwashina, T., Kilani, R.T. and Ghahary, A., Immune cell proliferation is suppressed by the interferon-gamma-induced indoleamine 2,3-dioxygenase expression of fibroblasts populated in collagen gel (FPCG), J Cell Biochem,. 90(1): 206-217, 2003.
[5]	Haniffa, M.A., Wang, X.N., Holtick, U., Rae, M., Isaacs, J.D., Dickinson, A.M., Hilken, C.M. and Collin, M.P., Adult human fibroblasts are potent immunoregulatory cells and functionally equivalent to mesenchymal stem cells, J Immunol., 179(3): 1595-1604, 2007.
[6]	Fehrer, C., Brunauer, R., Laschober, G., Unterluggauer, H., Reitinger, S., Kloss, F., Gully, C., Gassner, R. and Lepperdinger, G., Reduced oxygen tension attenuates differentiation capacity of human mesenchymal stem cells and prolongs their lifespan, Aging Cell. 6(6): 745-757, 2007.
[7]	Buravkova, L.B., Grinakovskaya, O.S., Andreeva, E.P., Zhambalova, A.P. and Kozionova, M.P., Characteristics of human lipoaspirate-isolated mesenchymal stromal cells cultivated under a lower oxygen tension, Cell and Tissue Biology., 3(1): 23-28, 2009.
[8]	Zuk, P., Zhu, M., Ashjian, P., De Ugarte, D.A., Huang, J.I., Mizuno, H., Alfonso, Z.C., Fraser, J.K., Benhaim, P. and Hedrick, M.H., Human adipose tissue is a source of multipotent stem cell , Mol Biol Cell., 13(12): 4279-4295, 2002.
[9]	Friedenstein, A.J., Gorskaja, J.F. Sep Exp Hematol 1976, 4:267-274Kulagina, N.N., Fibroblast precursors in normal and irradiated mouse hematopoietic organs, Exp Hematol., 4(5): 267-274, 1976.
[10]	Ishii, M., Koike, C., Igarashi, A., Yamanaka, K., Pan, H., Higashi, Y., Kawaguchi, H., Sugiyama, M., Kamata, N., Iwata, T., Matsubara, T., Nakamura, K., Kurihara, H., Tsuji, K. and Kato., Y., Molecular markers distinguish bone marrow mesenchymal stem cells from fibroblasts, , 332(1): 297-303, 2005.
[11]	Bae, S., Ahn, J.H., Park, C.W., Son, H.K., Kim, K.S., Lim, N.K., Jeon, C.J. and Kim, H., Gene and microRNA expression signatures of human mesenchymal stromal cells in comparison to fibroblasts , Cell Tissue Res., 335(3).565-573, 2009.
[12]	Sabatini, F., Petecchia, L., Tavian, M., Jodon de Villeroché, V., Rossi, G.A. and Brouty-Boyé, D., Human bronchial fibroblasts exhibit a mesenchymal stem cell phenotype and multilineage differentiating potentialities, Lab Invest. 85: 962-971, 2005.
[13]	Filer, A., Parsonage, G., Smith, E., Osborne, C., Thomas, A.M., Curnow, S.J., Rainger, G.E., Raza, K., Nash, G.B., Lord, J., Salmon, M. and Buckley, C.D., Differential survival of leukocyte subsets mediated by synovial, bone marrow, and skin fibroblasts, Arthritis Rheum. 54(7): 2096-2108, 2006.
[14]	Horwitz, E.M., Le Blanc, K., Dominici, M., Mueller, I., Slaper-Cortenbach, I., Marini, F.., Deans, R.J., Krause, D.S. and Keating, A., “Clarification of the nomenclature for MSC: The International Society for Cellular Therapy position statement,” Cytotherapy. 7.393-395. 2005.
[15]	Haniffa, M.A., Collin, M.P., Buckley, C.D. and Dazzi, F., “Mesenchymal stem cells: the fibroblasts' new clothes?,” Haematologica. 94.258-263. Feb.2009.
[16]	Potian, J.A., Aviv, H., Ponzio, N.M., Harrison, J.S. and Rameshwar, P., “Veto-like activity of mesenchymal stem cells: Functional discrimination between cellular responses to alloantigens and recall antigens,” J Immunol. 171.3426-3434. Oct.2003.
[17]	Harris, J., “Fibroblasts and their transformations: the connective-tissue cell family,” In Molecular Biology of the Cell. 3rd Edition. Edited by Alberts, B., Bra,y D., Lewis, J., Raff, M., Roberts, K. and Watson, .JD. New York: Garland Publishing; 1994:1179-1193.
[18]	Takahashi, K. and Yamanaka S., “Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors,” Cell. 126.663- 676. Aug.2006.
[19]	Kundig, T.M., Bachmann, M.F., DiPaolo, C., Simard, J.J., Battegay, M., Lother, H., Gessner, A., Kuhlcke, K., Ohashi, P.S., Hengartner, H. and Zinkernagel, R.M., “Fibroblasts as efficient antigen-presenting cells in lymphoid organs,” Science. 268.1343-1347. Jun.1995.
[20]	Korn, J.H., “Modulation of lymphocyte mitogen responses by cocultured fibroblasts,” Cell Immunol. 63.374-384. Sep.1981.
[21]	Suva, D., Passweg, J., Arnaudeau, S., Hoffmeyer, P. and Kindler, V., “In vitro activated human T lymphocytes very efficiently attach to allogenic multipotent mesenchymal stromal cells and transmigrate under them,” J Cell Physiol. 214.588-594. Mar.2008.
[22]	Quaedackers, M.E., Baan, C.C., Weimar, W. and Hoogduijn, M.J., “Cell contact interaction between adipose-derived stromal cells and allo-activated T lymphocytes,” Eur J Immunol. 39.3436-3446. Dec.2009.
[23]	Andreeva, E.R., Gornostaeva, A.N., Andrianova, I.V., Grigorieva, O.G., Buravkov, S.V., Rylova, Y.V. and Buravkova, L.B., “Morphologic aspects of immune and multipotent mesenchymal stromal cells' interactions in vitro. In: Stem cells and regenerative medicine. Edited by Tkachk, V.A., Мoscow: Maks-press: 2011:131-145.
[24]	Dexter, T.M., “Stromal cell associated haemopoiesis,” J Cell Physiol. Suppl. 1.87-94. 1982.
[25]	Hofmeister, C.C., Zhang, J., Knight, K.L., Le, P. and Stiff PJ., “Ex vivo expansion of umbilical cord blood stem cells for transplantation: growing knowledge from the hematopoietic niche,” Bone Marrow Transplant. 39.11-23. Jan.2007.
[26]	Wagner, W., Wein, F., Seckinger, A., Frankhauser, M., Wirkner, U., Krause, U., Blake, J., Schwager, C., Eckstein, V., Ansorge, W. and Ho, A.D., “Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood,” Experimental Hematology. 33.1402-1416. Nov.2005.
[27]	Kobari, L., Dubart, A., Le Pesteur, F., Vainchenker, W. And Sainteny, F., “Hematopoietic-promoting activity of the murine stromal cell line MS-5 is not related to the expression of the major hematopoietic cytokines,” J Cell Physiol. 163.295-304. May.1995.
[28]	Aoyama, K., Oritani, K., Yokota, T., Ishikawa, J., Nishiura, T., Miyake, K., Kanakura, Y., Tomiyama, Y., Kincade, P.W. “Matsuzawa Y: Stromal cell CD9 regulates differentiation of hematopoietic stem/progenitor cells,” Blood. 93.2586-2594. Apr.1999.
[29]	Freund, D., Bauer, N., Boxberger, S., Feldmann, S., Streller, U., Ehninger, G., Werner, C., Bornhäuser, M., Oswald, J. and Corbeil, D., “Polarization of human hematopoietic progenitors during contact with multipotent mesenchymal stromal cells: effects on proliferation and clonogenicity,” Stem Cells Dev. 15.815-829. Dec.2006.
[30]	Wagner, W., Roderburg, C., Wein, F., Diehlmann, A., Frankhauser, M., Schubert, R., Eckstein, V. and Ho, A.D., “Molecular and secretory profiles of human mesenchymal stromal cells and their abilities to maintain primitive hematopoietic progenitors,” “Stem Cells. 25.2638-2647. Oct.2007.
[31]	Arai, F. and Suda, T., “Maintenance of quiescent hematopoietic stem cells in the osteoblastic niche,” Ann N Y Acad Sci. 1106.41-53. Jun.2007.
[32]	Walenda, T., Bork, S., Horn, P., Wein, F., Saffrich, R., Diehlmann, A., Eckstein, V., Ho, A.D. and Wagner, W., “Co-culture with mesenchymal stromal cells increases proliferation and maintenance of haematopoietic progenitor cells,” J Cell Mol Med. 14.337-350. Jan.2010.
[33]	Li, N., Feugier, P., Serrurrier, B., Latger-Cannard, V., Lesesve, J.F., Stoltz, J.F. and Eljaafari, A., “Human mesenchymal stem cells improve ex vivo expansion of adult human CD34+ peripheral blood progenitor cells and decrease their allostimulatory capacity,” Exp Hematol. 35.507-515. Mar.2007.
[34]	Sitkovsky, M. and Lukashev, D., “Regulation of immune cells by local tissue oxigen tension: HIF1alpha and adenosine receptors,” Nat Rev Immunol. 5.712-721. Sep.2005.
[35]	Roman, J., Rangasamy, T., Guo, J., Sugunan, S., Meednu, N., Packirisamy, G., Shimoda, L.A., Golding, A., Semenza, G. and Georas, S.N., “T-cell activation under hypoxic conditions enhances IFN-gamma secretion,” Am J Respir Cell Mol Biol..42.123-128. Jan.2010
[36]	Buravkova, L.B., Grigorieva, O.G., Andreeva, E.R., Andrianova, I.V. and Rylova, Y.V., “Subpopulation Composition and Activation of T Lymphocytes during Coculturing with Mesenchymal Stromal Cells in Medium with Different O2 Content,” Bull Exp Biol Med. 151.344-346. Jul.2011.
[37]	Grinakovskaya, O.S., Andreeva, E.R., Buravkova, L.B., Rylova, Y.V, and Kosovsky, G.Y., “ow level of O2 inhibits commitment of cultured mesenchymal stromal precursor cells from the adipose tissue in response to osteogenic stimuli,” Bull Exp Biol Med. 147.760-763. Jun.2009.
[38]	Ma, T., Grayson, W.L., Frohlich, M. and Vunjak-Novakovic G., “Hypoxia and stem cell-based engineering of mesenchymal tissues,” Biotechnol Prog. 25.32-42. Jan-Feb.2009.
[39]	Cipolleschi, M.G., Dellosbarba, P. and Olivotto M., “The role of hypoxia in the maintenance of hematopoietic stem-cells,” Blood. 2.2031-2037. Oct.1993.
[40]	Parmar, K., Mauch, P., Vergilio, J.A., Sackstein, R. and Down, J.D., “Distribution of hematopoietic stem cells in the bone marrow according to regional hypoxia,” Proc Natl Acad Sci USA. 104.5431-5436. Mar.2007.
[41]	Shima, H., Takubo, K., Iwasaki, H., Yoshihara, H., Gomei, Y., Hosokawa, K., Arai, F., Takahashi, T. and Suda, T., “Reconstitution activity of hypoxic cultured human cord blood CD34-positive cells in NOG mice,” Biochem Biophys Res Commun. 378.467-472. Jan.2009.
[42]	Ivanovic, Z., Hermitte, F., Brunet de , P., Dazey, B., Belloc, F., Lacombe, F., Vezon, G. and Praloran, V., “Simultaneous maintenance of human cord blood SCID-repopulating cells and expansion of committed progenitors at low O2 concentration (3%),” Stem Cells. 2.716-724. 2004.
[43]	Zhambalova, A.P., Darevskaya, A.N., Kabaeva, N.V., Romanov, Y.A., Buravkova, L.B., “Specific interaction of cultured human mesenchymal and hemopoietic stem cells under conditions of reduced oxygen content,” Bull Exp Biol Med. 147.525-30. Apr.2009.