, Volume 70, Issue 1, pp 299–312 | Cite as

Interaction of allogeneic adipose tissue-derived stromal cells and unstimulated immune cells in vitro: the impact of cell-to-cell contact and hypoxia in the local milieu

  • Aleksandra N. Gornostaeva
  • Elena R. Andreeva
  • Polina I. Bobyleva
  • Ludmila B. Buravkova
Original Article


Multipotent mesenchymal stem cells (MSCs) are an attractive tool for cell therapy and regenerative medicine. Being applied in vivo, allogeneic MSCs are faced with both activated and unstimulated immune cells. The effects of MSCs on activated immune cells are well described and are mainly suppressive. Less is known about the interaction of MSCs with unstimulated immune cells. We evaluated the contribution of tissue-related O2 level (“physiological” hypoxia—5% O2) and cell-to-cell contact to the interaction between allogeneic adipose tissue-derived MSCs (ASCs) and unstimulated peripheral blood mononuclear cells (PBMCs). Under both O2 levels, ASCs affected the immune response by elevating the proportion of CD69+ T cells and modifying the functional activity of unstimulated PBMCs, providing a significant reduction of ROS level and activation of lysosome compartment. “Physiological” hypoxia partially attenuated the ASC modulation of PBMC function, reducing CD69+ cell activation and more significantly supressing ROS. In direct co-culture, the ASC effects were more pronounced. PBMC viability was preferentially maintained, and the lymphocyte subset ratio was altered in favour of B cells. Our findings demonstrate that allogeneic ASCs do not enhance the activation of unstimulated immune cells and can provide supportive functions. The “hypoxic” phenotype of ASCs may be more “desirable” for the interaction with allogeneic immune cells that may be required in cell therapy protocols.


MSC Lymphocytes Immunosuppression Cell-to-cell interaction Hypoxia Immune response 



Multipotent mesenchymal stem cells


Adipose stromal cells


Peripheral blood mononuclear cells


Mixed lymphocyte reaction


5,6-carboxyfluorescein diacetate succinimidyl ester



The study was funded by Programme of Presidium of Russian Academy of Sciences “Integrative physiology” and Grant of the President of the Russian Federation SP-3502.2015.4.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Ankrum JA, Ong JF, Karp JM (2014) Mesenchymal stem cells: immune evasive, not immune privileged. Nat Biotechnol 32:252–260. doi: 10.1038/nbt.2816 CrossRefGoogle Scholar
  2. Benvenuto F, Ferrari S, Gerdoni E, Gualandi F, Frassoni F, Pistoia V, Mancardi G, Uccelli A (2007) Human mesenchymal stem cells promote survival of T cells in a quiescent state. Stem Cells 25:1753–1760. doi: 10.1634/stemcells.2007-0068 CrossRefGoogle Scholar
  3. Bobyleva PI, Andreeva ER, Gornostaeva AN, Buravkova LB (2016) Tissue-related hypoxia attenuates proinflammatory effects of allogeneic pbMCS on adipose-derived stromal cells in vitro. Stem Cells Int 2016:4726267. doi:  10.1155/2016/4726267
  4. Bourin P, Bunnell BA, Casteilla L, Dominici M, Katz AJ, March KL, Redl H, Rubin JP, Yoshimura K, Gimble JM (2013) Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells: a joint statement of the International Federation for Adipose Therapeutics and Science (IFATS) and the International Society for Cellular Therapy (ISCT). Cytotherapy 15:641–648. doi: 10.1016/j.jcyt.2013.02.006 CrossRefGoogle Scholar
  5. Buravkova LB, Grinakovskaia OS, Andreeva EP, Zhambalova AP, Kozionova MP (2009) Characteristics of human lipoaspirate-isolated mesenchymal stromal cells cultivated under a lower oxygen tension. Cell Tiss Biol 3:23. doi: 10.1134/S1990519X09010039 CrossRefGoogle Scholar
  6. Buravkova LB, Rylova YV, Andreeva ER, Kulikov AV, Pogodina MV, Zhivotovsky B, Gogvadze V (2013) Low ATP level is sufficient to maintain the uncommitted state of multipotent mesenchymal stem cells. Biochim Biophys Acta 1830:4418–4425. doi: 10.1016/j.bbagen.2013.05.029 CrossRefGoogle Scholar
  7. Caldwell CC, Kojima H, Lukashev D, Armstrong J, Farber M, Apasov SG, Sitkovsky MV (2001) Differential effects of physiologically relevant hypoxic conditions on T lymphocyte development and effector functions. J Immunol 167:6140–6149. doi: 10.4049/jimmunol.167.11.6140 CrossRefGoogle Scholar
  8. Caplan AI, Sorrell JM (2015) The MSC curtain that stops the immune system. Immunol Lett 168:136–139. doi: 10.1016/j.imlet.2015.06.005 CrossRefGoogle Scholar
  9. Cappellesso-Fleury S, Puissant-Lubrano B, Apoil PA, Titeux M, Winterton P, Casteilla L, Bourin P, Blancher A (2010) Human fibroblasts share immunosuppressive properties with bone marrow mesenchymal stem cells. J Clin Immunol 30:607–619. doi: 10.1007/s10875-010-9415-4 CrossRefGoogle Scholar
  10. Chan JL, Tang KC, Patel AP, Bonilla LM, Pierobon N, Ponzio NM, Rameshwar P (2006) Antigen-presenting property of mesenchymal stem cells occurs during a narrow window at low levels of interferon-γ. Blood 107:4817–4824. doi: 10.1182/blood-2006-01-0057 CrossRefGoogle Scholar
  11. Chung YM, Kim JS, Yoo YD (2006) A novel protein, Romo1, induces ROS production in the mitochondria. Biochem Biophys Res Commun 347:649–655. doi: 10.1016/j.bbrc.2006.06.140 CrossRefGoogle Scholar
  12. Cipolleschi MG, Dello Sbarba P, Olivotto M (1993) The role of hypoxia in the maintenance of haematopoietic stem cells. Blood 82:2031–2037Google Scholar
  13. Conforti L, Petrovic M, Mohammad D, Lee S, Ma Q, Barone S, Filipovich AH (2003) Hypoxia regulates expression and activity of Kv1.3 channels in T lymphocytes: a possible role in T cell proliferation. J Immunol 170:695–702. doi: 10.4049/jimmunol.170.2.695 CrossRefGoogle Scholar
  14. Consentius C, Reinke P, Volk HD (2015) Immunogenicity of allogeneic mesenchymal stromal cells: what has been seen in vitro and in vivo? Regen Med 10:305–315. doi: 10.2217/rme.15.14 CrossRefGoogle Scholar
  15. Crop M, Baan CC, Korevaar SS, Ijzermans JN, Weimar W, Hoogduijn MJ (2010) Human adipose tissue-derived mesenchymal stem cells induce explosive T-cell proliferation. Stem Cells Dev 19:1843–1853. doi: 10.1089/scd.2009.0368 CrossRefGoogle Scholar
  16. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Dj Prockop, Horwitz E (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The international Society for Cellular Therapy position statement. Cytotherapy 8:315–317CrossRefGoogle Scholar
  17. Fehrer C, Brunauer R, Laschober G, Unterluggauer H, Reitinger S, Kloss F, Gülly C, Gassner R, Lepperdinger G (2007) Reduced oxygen tension attenuates differentiation capacity of human mesenchymal stem cells and prolongs their lifespan. Aging Cell 6:745–757. doi: 10.1111/j.1474-9726.2007.00336.x CrossRefGoogle Scholar
  18. Gonzalo-Daganzo R, Regidor C, Martín-Donaire T, Rico MA, Bautista G, Krsnik I, Forés R, Ojeda E, Sanjuán I, García-Marco JA, Navarro B, Gil S, Sánchez R, Panadero N, Gutiérrez Y, García-Berciano M, Pérez N, Millán I, Cabrera R, Fernández MN (2009) Results of a pilot study on the use of third-party donor mesenchymal stromal cells in cord blood transplantation in adults. Cytotherapy 11:278–288. doi: 10.1080/14653240902807018 CrossRefGoogle Scholar
  19. Gornostaeva AN, Andreeva ER, Buravkova LB (2013) Human MMSC immunosuppressive activity at low oxygen tension: direct cell-to-cell contacts and paracrine regulation. Hum Physiol 39:136. doi: 10.1134/S0362119713020059 CrossRefGoogle Scholar
  20. Gornostaeva AN, Andreeva ER, Buravkova LB (2016) Factors governing the immunosuppressive effects of multipotent mesenchymal stromal cells in vitro. Cytotechnology 68:565–577. doi: 10.1007/s10616-015-9906-5 CrossRefGoogle Scholar
  21. Grayson WL, Zhao F, Bunnell B, Ma T (2007) Hypoxia enhances proliferation and tissue formation of human mesenchymal stem cells. Biochem Biophys Res Commun 358(3):948–953. doi: 10.1016/j.bbrc.2007.05.054 CrossRefGoogle Scholar
  22. Kaplan JM, Youd ME, Lodie TA (2011) Immunomodulatory activity of mesenchymal stem cells. Curr Stem Cell Res Ther 6:297–316. doi: 10.2174/157488811797904353 CrossRefGoogle Scholar
  23. Kassem M, Kristiansen M, Abdallah BM (2004) Mesenchymal stem cells: cell biology and potential use in therapy. Basic Clin Pharmacol Toxicol 95:209–214. doi: 10.1111/j.1742-7843.2004.pto950502.x CrossRefGoogle Scholar
  24. Krieger JA, Landsiedel JC, Lawrence DA (1996) Differential in vitro effects of physiological and atmospheric oxygen tension on normal human peripheral blood mononuclear cell proliferation, cytokine and immunoglobulin production. Int J Immunopharmacol 18:545–552. doi: 10.1016/S0192-0561(96)00057-4 CrossRefGoogle Scholar
  25. Kronsteiner B, Wolbank S, Peterbauer A, Hackl C, Redl H, van Griensven M, Gabriel C (2011) Human mesenchymal stem cells from adipose tissue and amnion influence T-cells depending on stimulation method and presence of other immune cells. Stem Cells Dev 20:2115–2126. doi: 10.1089/scd.2011.0031 CrossRefGoogle Scholar
  26. Le Blanc K, Rasmusson I, Götherström C, Seidel C, Sundberg B, Sundin M, Rosendahl K, Tammik C, Ringdén O (2004) Mesenchymal stem cells inhibit the expression of CD25 (Interleukin-2 Receptor) and CD38 on phytohaemagglutinin-activated lymphocytes. Scand J Immunol 60:307–315. doi: 10.1111/j.0300-9475.2004.01483.x CrossRefGoogle Scholar
  27. Le Blanc K, Frassoni F, Ball L, Locatelli F, Roelofs H, Lewis I, Lanino E, Sundberg B, Bernardo ME, Remberger M, Dini G, Egeler RM, Bacigalupo A, Fibbe W, Ringdén O, Developmental Committee of the European Group for Blood and Marrow Transplantation (2008) Mesenchymal stem cells for treatment of steroid-resistant, severe, acute graft-versus-host disease: a phase II study. Lancet 371:1579–1586. doi: 10.1016/S0140-6736(08)60690-X CrossRefGoogle Scholar
  28. Lee SB, Kim JJ, Kim TW, Kim BS, Lee MS, Yoo YD (2010) Serum deprivation-induced reactive oxygen species production is mediated by Romo1. Apoptosis 15:204–218. doi: 10.1007/s10495-009-0411-1 CrossRefGoogle Scholar
  29. Madrigal M, Rao KS, Riordan NH (2014) A review of therapeutic effects of mesenchymal stem cell secretions and induction of secretory modification by different culture methods. J Transpl Med 12:260. doi: 10.1186/s12967-014-0260-8 CrossRefGoogle Scholar
  30. Magin AS, Korfer NR, Partenheimer H, Lange C, Zander A, Noll T (2009) Primary cells as feeder cells for co-culture expansion of human hematopoetic stem cells from umbilical cord blood—a comparative study. Stem Cells Dev 18:173–186. doi: 10.1089/scd.2007.0273 CrossRefGoogle Scholar
  31. Malladi P, Xu Y, Chiou M, Giaccia AJ, Longaker MT (2006) Effect of reduced oxygen tension on chondrogenesis and osteogenesis in adipose-derived mesenchymal cells. Am J Physiol Cell Physiol 290:1139–1146. doi: 10.1152/ajpcell.00415.2005 CrossRefGoogle Scholar
  32. Murabayashi D, Mochizuki M, Tamaki Y, Nakahara T (2017) Practical methods for handling human periodontal ligament stem cells in serum-free and serum-containing culture conditions under hypoxia: implications for regenerative medicine. Hum Cell 30:169–180. doi: 10.1007/s13577-017-0161-2 CrossRefGoogle Scholar
  33. Najar M, Raicevic G, Fayyad-Kazan H, Bron D, Toungouz M, Lagneaux L (2016) Mesenchymal stromal cells and immunomodulation: a gathering of regulatory immune cells. Cytotherapy 18:160–171. doi: 10.1016/j.jcyt.2015.10.011 CrossRefGoogle Scholar
  34. Nauta AJ, Fibbe WE (2007) Immunomodulatory properties of mesenchymal stromal cells. Blood 110:3499–3506. doi: 10.1182/blood-2007-02-069716 CrossRefGoogle Scholar
  35. Nekanti U, Dastidar S, Venugopal P, Totey S, Ta M (2010) Increased proliferation and analysis of differential gene expression in human Wharton’s jelly-derived mesenchymal stromal cells under hypoxia. Int J Biol Sci 6:499–512. doi: 10.7150/ijbs.6.499 CrossRefGoogle Scholar
  36. Parish CR (1999) Fluorescent dyes for lymphocyte migration and proliferation studies. Immunol Cell Biol 77:499–508. doi: 10.1046/j.1440-1711.1999.00877.x CrossRefGoogle Scholar
  37. Puissant B, Barreau C, Bourin P, Clavel C, Corre J, Bousquet C, Taureau C, Cousin B, Abbal M, Laharrague P, Penicaud L, Casteilla L, Blancher A (2005) Immunomodulatory effect of human adipose tissue-derived adult stem cells: comparison with bone marrow mesenchymal stem cells. Br J Haematol 129:118–129. doi: 10.1111/j.1365-2141.2005.05409.x CrossRefGoogle Scholar
  38. Ren G, Zhang L, Zhao X, Xu G, Zhang Y, Roberts AI, Zhao RC, Shi Y (2008) Mesenchymal stem cell-mediated immunosuppression occurs via concerted action of chemokines and nitric oxide. Cell Stem Cell 2:141–150. doi: 10.1016/j.stem.2007.11.014 CrossRefGoogle Scholar
  39. Sade H, Sarin A (2004) Reactive oxygen species regulate quiescent T-cell apoptosis via the BH3-only proapoptotic protein BIM. Cell Death Differ 11:416–423. doi: 10.1038/sj.cdd.4401347 CrossRefGoogle Scholar
  40. Sisakhtnezhad S, Alimoradi E, Akrami H (2017) External factors influencing mesenchymal stem cell fate in vitro. Eur J Cell Biol 96:13–33. doi: 10.1016/j.ejcb.2016.11.003 CrossRefGoogle Scholar
  41. Sitkovsky M, Lukashev D (2005) Regulation of immune cells by local tissue oxygen tension: HIF1α and adenosine receptors. Nat Rev Immunol 5:712–721. doi: 10.1038/nri1685 CrossRefGoogle Scholar
  42. Sun J, Zhang Y, Yang M, Zhang Y, Xie Q, Li Z, Dong Z, Yang Y, Deng B, Feng A, Hu W, Mao H, Qu X (2010) Hypoxia induces T-cell apoptosis by inhibiting chemokine C receptor 7 expression: the role of adenosine receptor A(2). Cell Mol Immunol 7:77–82. doi: 10.1038/cmi.2009.105 CrossRefGoogle Scholar
  43. Suva D, Passweg J, Arnaudeau S, Hoffmeyer P, Kindler V (2008) 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. doi: 10.1002/jcp.21244 CrossRefGoogle Scholar
  44. Tremp M, Meyer Zu Schwabedissen M, Kappos EA, Engels PE, Fischmann A, Scherberich A, Schaefer DJ, Kalbermatten DF (2015) The regeneration potential after human and autologous stem cell transplantation in a rat sciatic nerve injury model can be monitored by MRI. Cell Transpl 24:203–211CrossRefGoogle Scholar
  45. Wang LT, Ting CH, Yen ML, Liu KJ, Sytwu HK, Wu KK, Yen BL (2016) Human mesenchymal stem cells (MSCs) for treatment towards immune- and inflammation-mediated diseases: review of current clinical trials. J Biomed Sci 23:76. doi: 10.1186/s12929-016-0289-5 CrossRefGoogle Scholar
  46. Yang SH, Park MJ, Yoon IH, Kim SY, Hong SH, Shin JY, Nam HY, Kim YH, Kim B, Park CG (2009) Soluble mediators from mesenchymal stem cells supress T cell proliferation by inducing IL-10. Exp Mol Med 41:315–324. doi: 10.3858/emm.2009.41.5.035 CrossRefGoogle Scholar
  47. Yoo KH, Jang IK, Lee MW (2009) Comparison of immunomodulatory properties of mesenchymal stem cells derived from adult human tissues. Cell Immunol 259:150–156. doi: 10.1016/j.cellimm.2009.06.010 CrossRefGoogle Scholar
  48. Yu J, Zhang L (2008) PUMA, a potent killer with or without p53. Oncogene Suppl 1:S71–S83. doi: 10.1038/onc.2009.45 CrossRefGoogle Scholar
  49. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7:211–218. doi: 10.1089/107632701300062859 CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  1. 1.Cell Physiology Laboratory, Institute of Biomedical ProblemsRussian Academy of SciencesMoscowRussia

Personalised recommendations