Skip to main content
Download PDF
  • Research article
  • Published:

Mesenchymal stem cells promote a primitive phenotype CD34+c-kit+ in human cord blood-derived hematopoietic stem cells during ex vivo expansion

Cellular & Molecular Biology Letters volume 18, pages 11–33 (2013)Cite this article

Abstract

The purpose of this study was to evaluate the influence of bone marrow-mesenchymal stem cells (BM-MSC) and exogenously added cytokines on the proliferation, primitive cell subpopulation maintenance (including the c-kit+ marker) and clonogenic capacity of hematopoietic stem cells (HSC). BM-MSC were collected from volunteer donors, isolated and characterized. Umbilical cord blood (UCB) samples were collected from healthy full-term deliveries. UCB-CD34+ cells were cultured in the presence or absence of BM-MSC and/or cytokines for 3 and 7 days. CD34+ cell proliferation was evaluated using the CSFE method and cell phenotype was determined by CD34, c-kit, CD33, CD38, HLA-DR, cyCD22 and cyCD3 detection. Cell clonogenic ability was also assessed. Exogenously added SCF, TPO and FLT3L increasedCD34+ cell proliferation in the presence or absence of BM-MSC, but with concomitant cell differentiation. Without any added cytokines, BM-MSC are able to increase the percentage of primitive progenitors as evaluated by c-kit expression and CFU-GEMM increase. Interestingly, this latter effect was dependent on both cell-cell interactions and secreted factors. A 7-day co-culture period will be optimal for obtaining an increased primitive HSC level. Including c-kit as a marker for primitive phenotype evaluation has shown the relevance of BM-MSC and their secreted factors on UCB-HSC stemness function. This effect could be dissociated from that of the addition of exogenous cytokines, which induced cellular differentiation instead.

Abbreviations

BFU-E:

burst-forming unit erythroids

BM:

bone marrow

BMMSC:

bone marrow mesenchymal stem cells

CFSE:

carboxyfluorescein diacetate succinimidyl ester

CFU-E:

colony-forming unit erythroids

CFU-GEMM:

colonyforming unit-granulocyte, erythrocyte, monocyte, megakaryocyte

CFU-GM:

colonyforming unit-granulocyte, monocyte, macrophage

Flt-3L:

FMS-like tyrosine kinase 3 ligand

FRS:

frozen supernatant

FS:

fresh supernatant

HC:

hematopoietic cytokines

HM:

hematopoietic mesenchymal

HMC:

hematopoietic-mesenchymal cytokines

HSC:

hematopoietic stem cells

LTC-IC:

long-term culture-initiating cell

MACS:

magneticactivated cell sorting

MFI:

medium fluorescence intensity

MSC:

mesenchymal stem cells

SCF:

stem cell factor

TPO:

thrombopoietin

UCB:

umbilical cord blood

UCBCD34+:

umbilical cord blood-hematopoietic stem cell

References

  1. Rubinstein, P. Cord blood banking for clinical transplantation. Bone Marrow Transplant. 44 (2009) 635–642.

    Article  PubMed  CAS  Google Scholar 

  2. Robinson, S., Niu, T., de Lima, M., Ng, J., Yang, H., McMannis, J., Karandish, S., Sadeghi, T., Fu, P., del Angel, M., O’Connor, S., Champlin, R. and Shpall, E. Ex vivo expansion of umbilical cord blood. Cytotherapy 7 (2005) 243–250.

    Article  PubMed  CAS  Google Scholar 

  3. Cohena, Y. and Nagler, A. Hematopoietic stem-cell transplantation using umbilical-cord blood. Leuk. Lymphoma 44 (2003) 1287–1299.

    Article  PubMed  Google Scholar 

  4. Levac, K., Karanu, F. and Bhatia, M. Identification of growth factor conditions that reduce ex vivo cord blood progenitor expansion but do not alter human repopulating cell function in vivo. Haematologica 90 (2005) 166–172.

    PubMed  CAS  Google Scholar 

  5. Murray, L.J., Young, J.C., Osborne, L.J., Luens, K.M., Scollay, R. and Hill, B.L. Thrombopoietin, flt3, and kit ligands together suppress apoptosis of human mobilized CD34+ cells and recruit primitive CD34+ Thy-1+ cells into rapid division. Exp. Hematol. 27 (1999) 1019–1028.

    Article  PubMed  CAS  Google Scholar 

  6. Robinson, S.N., Ng, J., Niu, T., Yang, H., McMannis, J.D., Karandish, S., Kaur, I., Fu, P., Del Angel, M., Messinger, R., Flagge, F., de Lima, M., Decker, W., Xing, D., Champlin, R. and Shpall, E.J. Superior ex vivo cord blood expansion following co-culture with bone marrow-derived mesenchymal stem cells. Bone Marrow Transplant. 37 (2006) 359–366.

    Article  PubMed  CAS  Google Scholar 

  7. Glimm, H., Oh, I.H. and Eaves, C.J. Human hematopoietic stem cells stimulated to proliferate in vitro lose engraftment potential during their S/G(2)/M transit and do not reenter G(0). Blood 96 (2000) 4185–4193.

    PubMed  CAS  Google Scholar 

  8. Zhai, Q.L., Qiu, L.G., Li, Q., Meng, H.X., Han, J.L., Herzig, R.H. and Han, Z.C. Short-term ex vivo expansion sustains the homing-related properties of umbilical cord blood hematopoietic stem and progenitor cells. Haematologica 89 (2004) 265–273.

    PubMed  CAS  Google Scholar 

  9. Liu, B., Buckley, S.M., Lewis, I.D., Goldman, A.I., Wagner, J.E. and van der Loo, J.C. Homing defect of cultured human hematopoietic cells in the NOD/SCID mouse is mediated by Fas/CD95. Exp. Hematol. 31 (2003) 824–832.

    Article  PubMed  CAS  Google Scholar 

  10. Domen, J., Cheshier, S.H. and Weissman, I.L. The role of apoptosis in the regulation of hematopoietic stem cells: Overexpression of Bcl-2 increases both their number and repopulation potential. J. Exp. Med. 191 (2000) 253–264.

    Article  PubMed  CAS  Google Scholar 

  11. Wang, L.S., Liu, H.J., Xia, Z.B., Broxmeyer, H.E. and Lu, L. Expression and activation of caspase-3/CPP32 in CD34(+) cord blood cells is linked to apoptosis after growth factor withdrawal. Exp. Hematol. 28 (2000) 907–915.

    Article  PubMed  CAS  Google Scholar 

  12. Schofield, R. The relationship between the spleen colony-forming cell and the haemopoietic stem cell. Blood Cells 4 (1978) 7–25.

    PubMed  CAS  Google Scholar 

  13. Punzel, M., Liu, D., Zhang, T., Eckstein, V., Miesala, K. and Ho, A.D. The symmetry of initial divisions of human hematopoietic progenitors is altered only by the cellular microenvironment. Exp. Hematol. 31 (2003) 339–347.

    Article  PubMed  Google Scholar 

  14. Zhang, J., Niu, C., Ye, L., Huang, H., He, X., Tong, W.G., Ross, J., Haug, J., Johnson, T., Feng, J.Q., Harris, S., Wiedemann, L.M., Mishina, Y. and Li, L. Identification of the haematopoietic stem cell niche and control of the niche size. Nature 425 (2003) 836–841.

    Article  PubMed  CAS  Google Scholar 

  15. Dazzi, F., Ramasamy, R., Glennie, S., Jones, S.P. and Roberts, I. The role of mesenchymal stem cells in haemopoiesis. Blood Rev. 20 (2006) 161–171.

    Article  PubMed  CAS  Google Scholar 

  16. Flores-Guzman, P., Flores-Figueroa, E., Montesinos, J.J., Martinez-Jaramillo, G., Fernandez-Sanchez, V., Valencia-Plata, I., Alarcon-Santos, G. and Mayani, H. Individual and combined effects of mesenchymal stromal cells and recombinant stimulatory cytokines on the in vitro growth of primitive hematopoietic cells from human umbilical cord blood. Cytotherapy 11 (2009) 886–896.

    Article  PubMed  CAS  Google Scholar 

  17. McNiece, I., Harrington, J., Turney, J., Kellner, J. and Shpall, E.J. Ex vivo expansion of cord blood mononuclear cells on mesenchymal stem cells. Cytotherapy 6 (2004) 311–317.

    Article  PubMed  CAS  Google Scholar 

  18. 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 (2010) 337–350.

    Article  PubMed  CAS  Google Scholar 

  19. Magin, A.S., Korfer, N.R., Partenheimer, H., Lange, C., Zander, A. and Noll, T. Primary cells as feeder cells for coculture expansion of human hematopoietic stem cells from umbilical cord blood—a comparative study. Stem Cells Dev. 18 (2009) 173–186.

    Article  PubMed  CAS  Google Scholar 

  20. da Silva, C.L., Goncalves, R., dos Santos, F., Andrade, P.Z., Almeida-Porada, G. and Cabral, J.M. Dynamic cell-cell interactions between cord blood haematopoietic progenitors and the cellular niche are essential for the expansion of CD34+, CD34+CD38- and early lymphoid CD7+ cells. J. Tissue Eng. Regen. Med. 4 (2010) 149–158.

    Article  PubMed  Google Scholar 

  21. 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 (2007) 2638–2647.

    Article  PubMed  CAS  Google Scholar 

  22. Sutherland, H.J., Eaves, C.J., Eaves, A.C., Dragowska, W. and Lansdorp, P.M. Characterization and partial purification of human marrow cells capable of initiating long-term hematopoiesis in vitro. Blood 74 (1989) 1563–1570.

    PubMed  CAS  Google Scholar 

  23. Hao, Q.L., Thiemann, F.T., Petersen, D., Smogorzewska, E.M. and Crooks, G.M. Extended long-term culture reveals a highly quiescent and primitive human hematopoietic progenitor population. Blood 88 (1996) 3306–3313.

    PubMed  CAS  Google Scholar 

  24. Novelli, E.M., Ramirez, M. and Civin, C.I. Biology of CD34+CD38-cells in lymphohematopoiesis. Leuk. Lymphoma 31 (1998) 285–293.

    PubMed  CAS  Google Scholar 

  25. Glimm, H., Eisterer, W., Lee, K., Cashman, J., Holyoake, T.L., Nicolini, F., Shultz, L.D., von Kalle, C. and Eaves, C.J. Previously undetected human hematopoietic cell populations with short-term repopulating activity selectively engraft NOD/SCID-beta2 microglobulin-null mice. J. Clin. Invest. 107 (2001) 199–206.

    Article  PubMed  CAS  Google Scholar 

  26. Manz, M.G., Miyamoto, T., Akashi, K. and Weissman, I.L. Prospective isolation of human clonogenic common myeloid progenitors. Proc. Natl. Acad. Sci. USA 99 (2002) 11872–11877.

    Article  PubMed  CAS  Google Scholar 

  27. Bhatia, M., Bonnet, D., Kapp, U., Wang, J.C., Murdoch, B. and Dick, J.E. Quantitative analysis reveals expansion of human hematopoietic repopulating cells after short-term ex vivo culture. J. Exp. Med. 186 (1997) 619–624.

    Article  PubMed  CAS  Google Scholar 

  28. van Lochem, E.G., van der Velden, V.H., Wind, H.K., te Marvelde, J.G., Westerdaal, N.A. and van Dongen, J.J. Immunophenotypic differentiation patterns of normal hematopoiesis in human bone marrow: reference patterns for age-related changes and disease-induced shifts. Cytometry B. Clin. Cytom. 60 (2004) 1–13.

    Article  PubMed  Google Scholar 

  29. Yarden, Y., Kuang, W.J., Yang-Feng, T., Coussens, L., Munemitsu, S., Dull, T.J., Chen, E., Schlessinger, J., Francke, U. and Ullrich, A. Human protooncogene c-kit: a new cell surface receptor tyrosine kinase for an unidentified ligand. EMBO J. 6 (1987) 3341–3351.

    PubMed  CAS  Google Scholar 

  30. Miettinen, M. and Lasota, J. KIT (CD117): a review on expression in normal and neoplastic tissues, and mutations and their clinicopathologic correlation. Appl. Immunohistochem. Mol. Morphol. 13 (2005) 205–220.

    Article  PubMed  CAS  Google Scholar 

  31. Strobl, H., Takimoto, M., Majdic, O., Hocker, P. and Knapp, W. Antigenic analysis of human haemopoietic progenitor cells expressing the growth factor receptor c-kit. Br. J. Haematol. 82 (1992) 287–294.

    Article  PubMed  CAS  Google Scholar 

  32. Edling, C.E. and Hallberg, B. c-Kit—a hematopoietic cell essential receptor tyrosine kinase. Int. J. Biochem. Cell Biol. 39 (2007) 1995–1998.

    Article  PubMed  CAS  Google Scholar 

  33. Simmons, P.J., Aylett, G.W., Niutta, S., To, L.B., Juttner, C.A. and Ashman, L.K. c-kit is expressed by primitive human hematopoietic cells that give rise to colony-forming cells in stroma-dependent or cytokine-supplemented culture. Exp. Hematol. 22 (1994) 157–165.

    PubMed  CAS  Google Scholar 

  34. Hoffman, R., Tong, J., Brandt, J., Traycoff, C., Bruno, E., McGuire, B.W., Gordon, M.S., McNiece, I. and Srour, E.F. The in vitro and in vivo effects of stem cell factor on human hematopoiesis. Stem Cells 11Suppl 2 (1993) 76–82.

    PubMed  CAS  Google Scholar 

  35. Matarraz, S., Lopez, A., Barrena, S., Fernandez, C., Jensen, E., Flores, J., Barcena, P., Rasillo, A., Sayagues, J.M., Sanchez, M.L., Hernandez-Campo, P., Hernandez Rivas, J.M., Salvador, C., Fernandez-Mosteirin, N., Giralt, M., Perdiguer, L. and Orfao, A. The immunophenotype of different immature, myeloid and B-cell lineage-committed CD34+ hematopoietic cells allows discrimination between normal/reactive and myelodysplastic syndrome precursors. Leukemia 22 (2008) 1175–1183.

    Article  PubMed  CAS  Google Scholar 

  36. Pittenger, M.F., Mackay, A.M., Beck, S.C., Jaiswal, R.K., Douglas, R., Mosca, J.D., Moorman, M.A., Simonetti, D.W., Craig, S. and Marshak, D.R. Multilineage potential of adult human mesenchymal stem cells. Science 284 (1999) 143–147.

    Article  PubMed  CAS  Google Scholar 

  37. Phillips, J.E., Gersbach, C.A., Wojtowicz, A.M. and Garcia, A.J. Glucocorticoid-induced osteogenesis is negatively regulated by Runx2/Cbfa1 serine phosphorylation. J. Cell Sci. 119 (2006) 581–591.

    Article  PubMed  CAS  Google Scholar 

  38. Rosen, E.D. The transcriptional basis of adipocyte development. Prostaglandins Leukot. Essent. Fatty Acids 73 (2005) 31–34.

    Article  PubMed  CAS  Google Scholar 

  39. Han, F., Gilbert, J.R., Harrison, G., Adams, C.S., Freeman, T., Tao, Z., Zaka, R., Liang, H., Williams, C., Tuan, R.S., Norton, P.A. and Hickok, N.J. Transforming growth factor-beta1 regulates fibronectin isoform expression and splicing factor SRp40 expression during ATDC5 chondrogenic maturation. Exp. Cell Res. 313 (2007) 1518–1532.

    Article  PubMed  CAS  Google Scholar 

  40. Chularojmontri, L. and Wattanapitayakul, S.K. Isolation and characterization of umbilical cord blood hematopoietic stem cells. J. Med. Assoc. Thai. 92Suppl 3 (2009) S88–94.

    PubMed  Google Scholar 

  41. Harvey, K. and Dzierzak, E. Cell-cell contact and anatomical compatibility in stromal cell-mediated HSC support during development. Stem Cells 22 (2004) 253–258.

    Article  PubMed  Google Scholar 

  42. Jang, Y.K., Jung, D.H., Jung, M.H., Kim, D.H., Yoo, K.H., Sung, K.W., Koo, H.H., Oh, W., Yang, Y.S. and Yang, S.E. Mesenchymal stem cells feeder layer from human umbilical cord blood for ex vivo expanded growth and proliferation of hematopoietic progenitor cells. Ann. Hematol. 85 (2006) 212–225.

    Article  PubMed  Google Scholar 

  43. Ogawa, M. Differentiation and proliferation of hematopoietic stem cells. Blood 81 (1993) 2844–2853.

    PubMed  CAS  Google Scholar 

  44. Lessard, J., Faubert, A. and Sauvageau, G. Genetic programs regulating HSC specification, maintenance and expansion. Oncogene 23 (2004) 7199–7209.

    Article  PubMed  CAS  Google Scholar 

  45. Yu, S., Cui, K., Jothi, R., Zhao, D.M., Jing, X., Zhao, K. and Xue, H.H. GABP controls a critical transcription regulatory module that is essential for maintenance and differentiation of hematopoietic stem/progenitor cells. Blood 117 (2011) 2166–2178.

    Article  PubMed  CAS  Google Scholar 

  46. Andrade, P.Z., dos Santos, F., Almeida-Porada, G., da Silva, C.L. and S. Cabral, J.M. Systematic delineation of optimal cytokine concentrations to expand hematopoietic stem/progenitor cells in co-culture with mesenchymal stem cells. Mol. Biosyst. 6 (2010) 1207–1215.

    Article  PubMed  CAS  Google Scholar 

  47. McKenna, H.J., de Vries, P., Brasel, K., Lyman, S.D. and Williams, D.E. Effect of flt3 ligand on the ex vivo expansion of human CD34+ hematopoietic progenitor cells. Blood 86 (1995) 3413–3420.

    PubMed  CAS  Google Scholar 

  48. Gabutti, V., Timeus, F., Ramenghi, U., Crescenzio, N., Marranca, D., Miniero, R., Cornaglia, G. and Bagnara, G.P. Expansion of cord blood progenitors and use for hemopoietic reconstitution. Stem Cells 11 Suppl 2 (1993) 105–112.

    Article  PubMed  Google Scholar 

  49. Rossmanith, T., Schroder, B., Bug, G., Muller, P., Klenner, T., Knaus, R., Hoelzer, D. and Ottmann, O.G. Interleukin 3 improves the ex vivo expansion of primitive human cord blood progenitor cells and maintains the engraftment potential of scid repopulating cells. Stem Cells 19 (2001) 313–320.

    Article  PubMed  CAS  Google Scholar 

  50. Tajima, S., Tsuji, K., Ebihara, Y., Sui, X., Tanaka, R., Muraoka, K., Yoshida, M., Yamada, K., Yasukawa, K., Taga, T., Kishimoto, T. and Nakahata, T. Analysis of interleukin 6 receptor and gp130 expressions and proliferative capability of human CD34+ cells. J. Exp. Med. 184 (1996) 1357–1364.

    Article  PubMed  CAS  Google Scholar 

  51. Schipper, L.F., Brand, A., Reniers, N.C., Melief, C.J., Willemze, R. and Fibbe, W.E. Effects of thrombopoietin on the proliferation and differentiation of primitive and mature haemopoietic progenitor cells in cord blood. Br. J. Haematol. 101 (1998) 425–435.

    Article  PubMed  CAS  Google Scholar 

  52. Ohmizono, Y., Sakabe, H., Kimura, T., Tanimukai, S., Matsumura, T., Miyazaki, H., Lyman, S.D. and Sonoda, Y. Thrombopoietin augments ex vivo expansion of human cord blood-derived hematopoietic progenitors in combination with stem cell factor and flt3 ligand. Leukemia 11 (1997) 524–530.

    Article  PubMed  CAS  Google Scholar 

  53. De Lima, M., McMannis, J.D., Saliba, R., Worth, L., Kebriaei, P., Popat, U., Qazilbash, M., Jones, R., Giralt, S., de Padua Silva, L., Cooper, L., Petropoulos, D., Lee, D., Kelly, S., Thall, P., Robinson, S., Khouri, I., Hosing, C., Korbling, M., Alousi, A., Rondon, G., Andersson, B.S., Nieto, Y., Ciurea, S., Komanduri, K., Champlin, R.E. and Shpall, E. Double Cord Blood Transplantation (CBT) with and without Ex-Vivo Expansion (EXP): A randomized, controlled study. ASH Annual Meeting Abstracts 112 (2008) 154.

    Google Scholar 

  54. Rabinowitz, J., Petros, W.P., Stuart, A.R. and Peters, W.P. Characterization of endogenous cytokine concentrations after high-dose chemotherapy with autologous bone marrow support. Blood 81 (1993) 2452–2459.

    PubMed  CAS  Google Scholar 

  55. Mortera-Blanco, T., Rende, M., Macedo, H., Farah, S., Bismarck, A., Mantalaris, A. and Panoskaltsis, N. Ex vivo mimicry of normal and abnormal human hematopoiesis. J. Vis. Exp. 62 (2012) 3654–3791

    PubMed  Google Scholar 

  56. Mortera-Blanco, T., Mantalaris, A., Bismarck, A., Aqel, N. and Panoskaltsis, N. Long-term cytokine-free expansion of cord blood mononuclear cells in three-dimensional scaffolds. Biomaterials 32 (2011) 9263–9270.

    Article  PubMed  CAS  Google Scholar 

  57. Pandey, A.C., Semon, J.A., Kaushal, D., O’sullivan, R.P., Glowacki, J., Gimble, J.M. and Bunnell, B.A. MicroRNA profiling reveals age-dependent differential expression of nuclear factor kappaB and mitogen-activated protein kinase in adipose and bone marrow-derived human mesenchymal stem cells. Stem Cell Res. Ther. 2 (2011) 49.

    Article  PubMed  CAS  Google Scholar 

  58. Kinzebach, S. and Bieback, K. Expansion of mesenchymal stem/stromal cells under xenogenic-free culture conditions. Adv. Biochem. Eng. Biotechnol. (2012) [Epub ahead of print]

  59. Bhatia, M., Wang, J.C., Kapp, U., Bonnet, D. and Dick, J.E. Purification of primitive human hematopoietic cells capable of repopulating immunedeficient mice. Proc. Natl. Acad. Sci. USA 94 (1997) 5320–5325.

    Article  PubMed  CAS  Google Scholar 

  60. Conneally, E., Cashman, J., Petzer, A. and Eaves, C. Expansion in vitro of transplantable human cord blood stem cells demonstrated using a quantitative assay of their lympho-myeloid repopulating activity in nonobese diabetic-scid/scid mice. Proc. Natl. Acad. Sci. USA 94 (1997) 9836–9841.

    Article  PubMed  CAS  Google Scholar 

  61. Hogan, C.J., Shpall, E.J. and Keller, G. Differential long-term and multilineage engraftment potential from subfractions of human CD34+ cord blood cells transplanted into NOD/SCID mice. Proc. Natl. Acad. Sci. USA 99 (2002) 413–418.

    Article  PubMed  CAS  Google Scholar 

  62. Butler, J.M., Nolan, D.J., Vertes, E.L., Varnum-Finney, B., Kobayashi, H., Hooper, A.T., Seandel, M., Shido, K., White, I.A., Kobayashi, M., Witte, L., May, C., Shawber, C., Kimura, Y., Kitajewski, J., Rosenwaks, Z., Bernstein, I.D. and Rafii, S. Endothelial cells are essential for the self-renewal and repopulation of Notch-dependent hematopoietic stem cells. Cell Stem Cell 6 (2010) 251–264.

    Article  PubMed  CAS  Google Scholar 

  63. Ding, L., Saunders, T.L., Enikolopov, G. and Morrison, S.J. Endothelial and perivascular cells maintain haematopoietic stem cells. Nature 481 (2012) 457–462.

    Article  PubMed  CAS  Google Scholar 

  64. Liao, J., Hammerick, K.E., Challen, G.A., Goodell, M.A., Kasper, F.K. and Mikos, A.G. Investigating the role of hematopoietic stem and progenitor cells in regulating the osteogenic differentiation of mesenchymal stem cells in vitro. J. Orthop. Res. 29 (2011) 1544–1553.

    Article  PubMed  CAS  Google Scholar 

  65. Ogawa, M., Matsuzaki, Y., Nishikawa, S., Hayashi, S., Kunisada, T., Sudo, T., Kina, T. and Nakauchi, H. Expression and function of c-kit in hemopoietic progenitor cells. J. Exp. Med. 174 (1991) 63–71.

    Article  PubMed  CAS  Google Scholar 

  66. Gorczyca, W., Sun, Z.Y., Cronin, W., Li, X., Mau, S. and Tugulea, S. Immunophenotypic pattern of myeloid populations by flow cytometry analysis. Methods Cell Biol. 103 (2011) 221–266.

    Article  PubMed  CAS  Google Scholar 

  67. Grundmann, F., Scheid, C., Braun, D., Zobel, C., Reuter, H., Schwinger, R.H. and Muller-Ehmsen, J. Differential increase of CD34, KDR/CD34, CD133/CD34 and CD117/CD34 positive cells in peripheral blood of patients with acute myocardial infarction. Clin. Res. Cardiol. 96 (2007) 621–627.

    Article  PubMed  CAS  Google Scholar 

  68. Li, T.S., Hamano, K., Nishida, M., Hayashi, M., Ito, H., Mikamo, A. and Matsuzaki, M. CD117+ stem cells play a key role in therapeutic angiogenesis induced by bone marrow cell implantation. Am. J. Physiol. Heart Circ. Physiol. 285 (2003) H931–937.

    PubMed  CAS  Google Scholar 

  69. Montani, D., Perros, F., Gambaryan, N., Girerd, B., Dorfmuller, P., Price, L.C., Huertas, A., Hammad, H., Lambrecht, B., Simonneau, G., Launay, J.M., Cohen-Kaminsky, S., and Humbert, M. C-kit-positive cells accumulate in remodeled vessels of idiopathic pulmonary arterial hypertension. Am. J. Respir. Crit. Care Med. 184 (2011) 116–123.

    Article  PubMed  Google Scholar 

  70. Gardner, J.P., Rosenzweig, M., Marks, D.F., Harper, D., Gaynor, K., Fallon, R.J., Wall, D.A., Johnson, R.P. and Scadden, D.T. T-lymphopoietic capacity of cord blood-derived CD34+ progenitor cells. Exp. Hematol. 26 (1998) 991–999.

    PubMed  CAS  Google Scholar 

  71. Robin, C., Bennaceur-Griscelli, A., Louache, F., Vainchenker, W. and Coulombel, L. Identification of human T-lymphoid progenitor cells in CD34+ CD38low and CD34+ CD38+ subsets of human cord blood and bone marrow cells using NOD-SCID fetal thymus organ cultures. Br. J. Haematol. 104 (1999) 809–819.

    Article  PubMed  CAS  Google Scholar 

  72. Fei, X.M., Wu, Y.J., Chang, Z., Miao, K.R., Tang, Y.H., Zhou, X.Y., Wang, L.X., Pan, Q.Q. and Wang, C.Y. Co-culture of cord blood CD34(+) cells with human BM mesenchymal stromal cells enhances short-term engraftment of cord blood cells in NOD/SCID mice. Cytotherapy 9 (2007) 338–347.

    Article  PubMed  CAS  Google Scholar 

  73. Nakamura, Y., Arai, F., Iwasaki, H., Hosokawa, K., Kobayashi, I., Gomei, Y., Matsumoto, Y., Yoshihara, H. and Suda, T. Isolation and characterization of endosteal niche cell populations that regulate hematopoietic stem cells. Blood 116 (2010) 1422–1432.

    Article  PubMed  CAS  Google Scholar 

  74. Mendez-Ferrer, S., Michurina, T.V., Ferraro, F., Mazloom, A.R., Macarthur, B.D., Lira, S.A., Scadden, D.T., Ma’ayan, A., Enikolopov, G.N. and Frenette, P.S. Mesenchymal and haematopoietic stem cells form a unique bone marrow niche. Nature 466 (2010) 829–834.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Grupo de Inmunobiología y Biología Celular, Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, Colombia

    Viviana M. Rodríguez-Pardo

  2. rupo de Fisiología Celular y Molecular, Facultad de Medicina, Universidad Nacional de Colombia, Bogotá, Colombia

    Jean Paul Vernot

  3. Universidad Nacional de Colombia, Carrera 30 No. 45-03 - Edificio 471, Bogotá D.C., Colombia

    Jean Paul Vernot

Authors
  1. Viviana M. Rodríguez-Pardo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jean Paul Vernot
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jean Paul Vernot.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rodríguez-Pardo, V.M., Vernot, J.P. Mesenchymal stem cells promote a primitive phenotype CD34+c-kit+ in human cord blood-derived hematopoietic stem cells during ex vivo expansion. Cell Mol Biol Lett 18, 11–33 (2013). https://doi.org/10.2478/s11658-012-0036-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2478/s11658-012-0036-1

Key words

Cellular & Molecular Biology Letters

ISSN: 1689-1392

Contact us