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Blood, 1 November 2001, Vol. 98, No. 9, pp. 2615-2625

PLENARY PAPER

Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells

Morayma Reyes, Troy Lund, Todd Lenvik, Dean Aguiar, Lisa Koodie, and Catherine M. Verfaillie

From the Stem Cell Institute, Department of Medicine, and Cancer Center, University of Minnesota Medical School, Minneapolis.

It is here reported that mesenchymal stem cells known to give rise to limb-bud mesoderm can, at the single-cell level, also differentiate into cells of visceral mesoderm and can be expanded extensively by means of clinically applicable methods. These cells were named mesodermal progenitor cells (MPCs). MPCs were selected by depleting bone marrow mononuclear cells from more than 30 healthy human donors of CD45+/glycophorin-A (GlyA)+ cells. Cells were cultured on fibronectin with epidermal growth factor and platelet-derived growth factor BB and 2% or less fetal calf serum. It was found that 1/5 × 103 CD45-GlyA- cells, or 1/106 bone marrow mononuclear cells, gave rise to clusters of small adherent cells. Cell-doubling time was 48 to 72 hours, and cells have been expanded in culture for more than 60 cell doublings. MPCs are CD34-, CD44low, CD45-, CD117 (cKit)-, class I-HLA-, and HLA-DR-. MPCs differentiated into cells of limb-bud mesoderm (osteoblasts, chondrocytes, adipocytes, stroma cells, and skeletal myoblasts) as well as visceral mesoderm (endothelial cells). Retroviral marking was used to definitively prove that single MPCs can differentiate into cells of limb bud and visceral mesoderm. Thus, MPCs that proliferate without obvious senescence under clinically applicable conditions and differentiate at the single-cell level not only into mesenchymal cells but also cells of visceral mesoderm may be an ideal source of stem cells for treatment of genetic or degenerative disorders affecting cells of mesodermal origin.

© 2001 by The American Society of Hematology.
 

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Stem CellsHome page
T. Tondreau, N. Meuleman, A. Delforge, M. Dejeneffe, R. Leroy, M. Massy, C. Mortier, D. Bron, and L. Lagneaux
Mesenchymal Stem Cells Derived from CD133-Positive Cells in Mobilized Peripheral Blood and Cord Blood: Proliferation, Oct4 Expression, and Plasticity
Stem Cells, September 1, 2005; 23(8): 1105 - 1112.
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Stem CellsHome page
S. M. Majka, M. A. Beutz, M. Hagen, A. A. Izzo, N. Voelkel, and K. M. Helm
Identification of Novel Resident Pulmonary Stem Cells: Form and Function of the Lung Side Population
Stem Cells, September 1, 2005; 23(8): 1073 - 1081.
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BloodHome page
V. Sordi, M. L. Malosio, F. Marchesi, A. Mercalli, R. Melzi, T. Giordano, N. Belmonte, G. Ferrari, B. E. Leone, F. Bertuzzi, et al.
Bone marrow mesenchymal stem cells express a restricted set of functionally active chemokine receptors capable of promoting migration to pancreatic islets
Blood, July 15, 2005; 106(2): 419 - 427.
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BloodHome page
Y. Sato, H. Araki, J. Kato, K. Nakamura, Y. Kawano, M. Kobune, T. Sato, K. Miyanishi, T. Takayama, M. Takahashi, et al.
Human mesenchymal stem cells xenografted directly to rat liver are differentiated into human hepatocytes without fusion
Blood, July 15, 2005; 106(2): 756 - 763.
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Ann. Thorac. Surg.Home page
Y. L. Tang, Q. Zhao, X. Qin, L. Shen, L. Cheng, J. Ge, and M. I. Phillips
Paracrine Action Enhances the Effects of Autologous Mesenchymal Stem Cell Transplantation on Vascular Regeneration in Rat Model of Myocardial Infarction
Ann. Thorac. Surg., July 1, 2005; 80(1): 229 - 237.
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Mol. Biol. CellHome page
B. M. Deasy, B. M. Gharaibeh, J. B. Pollett, M. M. Jones, M. A. Lucas, Y. Kanda, and J. Huard
Long-Term Self-Renewal of Postnatal Muscle-derived Stem Cells
Mol. Biol. Cell, July 1, 2005; 16(7): 3323 - 3333.
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JEMHome page
A.-M. Rodriguez, D. Pisani, C. A. Dechesne, C. Turc-Carel, J.-Y. Kurzenne, B. Wdziekonski, A. Villageois, C. Bagnis, J.-P. Breittmayer, H. Groux, et al.
Transplantation of a multipotent cell population from human adipose tissue induces dystrophin expression in the immunocompetent mdx mouse
J. Exp. Med., May 2, 2005; 201(9): 1397 - 1405.
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Stem CellsHome page
C. Moriscot, F. de Fraipont, M.-J. Richard, M. Marchand, P. Savatier, D. Bosco, M. Favrot, and P.-Y. Benhamou
Human Bone Marrow Mesenchymal Stem Cells Can Express Insulin and Key Transcription Factors of the Endocrine Pancreas Developmental Pathway upon Genetic and/or Microenvironmental Manipulation In Vitro
Stem Cells, April 1, 2005; 23(4): 594 - 603.
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BloodHome page
B. Fang, C. Zheng, L. Liao, Q. Han, Z. Sun, X. Jiang, and R. C. H. Zhao
Identification of human chronic myelogenous leukemia progenitor cells with hemangioblastic characteristics
Blood, April 1, 2005; 105(7): 2733 - 2740.
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BloodHome page
S. Glennie, I. Soeiro, P. J. Dyson, E. W.-F. Lam, and F. Dazzi
Bone marrow mesenchymal stem cells induce division arrest anergy of activated T cells
Blood, April 1, 2005; 105(7): 2821 - 2827.
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Stem CellsHome page
G.-R. Li, H. Sun, X. Deng, and C.-P. Lau
Characterization of Ionic Currents in Human Mesenchymal Stem Cells from Bone Marrow
Stem Cells, March 1, 2005; 23(3): 371 - 382.
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Stem CellsHome page
E. Steck, H. Bertram, R. Abel, B. Chen, A. Winter, and W. Richter
Induction of Intervertebral Disc-Like Cells From Adult Mesenchymal Stem Cells
Stem Cells, March 1, 2005; 23(3): 403 - 411.
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J. Am. Soc. Nephrol.Home page
M. Iwasaki, Y. Adachi, K. Minamino, Y. Suzuki, Y. Zhang, M. Okigaki, K. Nakano, Y. Koike, J. Wang, H. Mukaide, et al.
Mobilization of Bone Marrow Cells by G-CSF Rescues Mice from Cisplatin-Induced Renal Failure, and M-CSF Enhances the Effects of G-CSF
J. Am. Soc. Nephrol., March 1, 2005; 16(3): 658 - 666.
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BloodHome page
S. Beyth, Z. Borovsky, D. Mevorach, M. Liebergall, Z. Gazit, H. Aslan, E. Galun, and J. Rachmilewitz
Human mesenchymal stem cells alter antigen-presenting cell maturation and induce T-cell unresponsiveness
Blood, March 1, 2005; 105(5): 2214 - 2219.
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Stem CellsHome page
R. Sarugaser, D. Lickorish, D. Baksh, M. M. Hosseini, and J. E. Davies
Human Umbilical Cord Perivascular (HUCPV) Cells: A Source of Mesenchymal Progenitors
Stem Cells, February 1, 2005; 23(2): 220 - 229.
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Am. J. Pathol.Home page
B. Bussolati, S. Bruno, C. Grange, S. Buttiglieri, M. C. Deregibus, D. Cantino, and G. Camussi
Isolation of Renal Progenitor Cells from Adult Human Kidney
Am. J. Pathol., February 1, 2005; 166(2): 545 - 555.
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Cardiovasc ResHome page
S. Davani, F. Deschaseaux, D. Chalmers, P. Tiberghien, and J.-P. Kantelip
Can stem cells mend a broken heart?
Cardiovasc Res, February 1, 2005; 65(2): 305 - 316.
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Stem CellsHome page
R. A. Panepucci, J. L.C. Siufi, W. A. Silva Jr., R. Proto-Siquiera, L. Neder, M. Orellana, V. Rocha, D. T. Covas, and M. A. Zago
Comparison of Gene Expression of Umbilical Cord Vein and Bone Marrow-Derived Mesenchymal Stem Cells
Stem Cells, December 1, 2004; 22(7): 1263 - 1278.
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JRSMHome page
H T Hassan and M El-Sheemy
Adult bone-marrow stem cells and their potential in medicine
J R Soc Med, October 1, 2004; 97(10): 465 - 471.
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Am. J. Physiol. Cell Physiol.Home page
T. Asahara and A. Kawamoto
Endothelial progenitor cells for postnatal vasculogenesis
Am J Physiol Cell Physiol, September 1, 2004; 287(3): C572 - C579.
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Stem CellsHome page
Y. Fukuchi, H. Nakajima, D. Sugiyama, I. Hirose, T. Kitamura, and K. Tsuji
Human Placenta-Derived Cells Have Mesenchymal Stem/Progenitor Cell Potential
Stem Cells, September 1, 2004; 22(5): 649 - 658.
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Stem CellsHome page
M. A. Baxter, R. F. Wynn, S. N. Jowitt, J. E. Wraith, L. J. Fairbairn, and I. Bellantuono
Study of Telomere Length Reveals Rapid Aging of Human Marrow Stromal Cells following In Vitro Expansion
Stem Cells, September 1, 2004; 22(5): 675 - 682.
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J. Cell Sci.Home page
A. Hermann, R. Gastl, S. Liebau, M. O. Popa, J. Fiedler, B. O. Boehm, M. Maisel, H. Lerche, J. Schwarz, R. Brenner, et al.
Efficient generation of neural stem cell-like cells from adult human bone marrow stromal cells
J. Cell Sci., September 1, 2004; 117(19): 4411 - 4422.
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Circ. Res.Home page
T. Kinnaird, E. Stabile, M. S. Burnett, and S. E. Epstein
Bone Marrow-Derived Cells for Enhancing Collateral Development: Mechanisms, Animal Data, and Initial Clinical Experiences
Circ. Res., August 20, 2004; 95(4): 354 - 363.
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Am. J. Physiol. Heart Circ. Physiol.Home page
J. Liu, Q. Hu, Z. Wang, C. Xu, X. Wang, G. Gong, A. Mansoor, J. Lee, M. Hou, L. Zeng, et al.
Autologous stem cell transplantation for myocardial repair
Am J Physiol Heart Circ Physiol, August 1, 2004; 287(2): H501 - H511.
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JEMHome page
G. Kogler, S. Sensken, J. A. Airey, T. Trapp, M. Muschen, N. Feldhahn, S. Liedtke, R. V. Sorg, J. Fischer, C. Rosenbaum, et al.
A New Human Somatic Stem Cell from Placental Cord Blood with Intrinsic Pluripotent Differentiation Potential
J. Exp. Med., July 19, 2004; 200(2): 123 - 135.
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Circ. Res.Home page
M. F. Pittenger and B. J. Martin
Mesenchymal Stem Cells and Their Potential as Cardiac Therapeutics
Circ. Res., July 9, 2004; 95(1): 9 - 20.
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Stem CellsHome page
E. J. Gang, J. A. Jeong, S. H. Hong, S. H. Hwang, S. W. Kim, I. H. Yang, C. Ahn, H. Han, and H. Kim
Skeletal Myogenic Differentiation of Mesenchymal Stem Cells Isolated from Human Umbilical Cord Blood
Stem Cells, July 1, 2004; 22(4): 617 - 624.
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J. Cell Sci.Home page
G. D'Ippolito, S. Diabira, G. A. Howard, P. Menei, B. A. Roos, and P. C. Schiller
Marrow-isolated adult multilineage inducible (MIAMI) cells, a unique population of postnatal young and old human cells with extensive expansion and differentiation potential
J. Cell Sci., June 15, 2004; 117(14): 2971 - 2981.
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Toxicol SciHome page
J. C. Davila, G. G. Cezar, M. Thiede, S. Strom, T. Miki, and J. Trosko
Use and Application of Stem Cells in Toxicology
Toxicol. Sci., June 1, 2004; 79(2): 214 - 223.
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Stem CellsHome page
J. Oswald, S. Boxberger, B. Jorgensen, S. Feldmann, G. Ehninger, M. Bornhauser, and C. Werner
Mesenchymal Stem Cells Can Be Differentiated Into Endothelial Cells In Vitro
Stem Cells, May 1, 2004; 22(3): 377 - 384.
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Stem CellsHome page
M. Murga, L. Yao, and G. Tosato
Derivation of Endothelial Cells from CD34- Umbilical Cord Blood
Stem Cells, May 1, 2004; 22(3): 385 - 395.
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Stem CellsHome page
S. Neuss, E. Becher, M. Woltje, L. Tietze, and W. Jahnen-Dechent
Functional Expression of HGF and HGF Receptor/c-met in Adult Human Mesenchymal Stem Cells Suggests a Role in Cell Mobilization, Tissue Repair, and Wound Healing
Stem Cells, May 1, 2004; 22(3): 405 - 414.
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BloodHome page
M. Bensidhoum, A. Chapel, S. Francois, C. Demarquay, C. Mazurier, L. Fouillard, S. Bouchet, J. M. Bertho, P. Gourmelon, J. Aigueperse, et al.
Homing of in vitro expanded Stro-1- or Stro-1+ human mesenchymal stem cells into the NOD/SCID mouse and their role in supporting human CD34 cell engraftment
Blood, May 1, 2004; 103(9): 3313 - 3319.
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BloodHome page
M. del Carmen Rodriguez, A. Bernad, and M. Aracil
Interleukin-6 deficiency affects bone marrow stromal precursors, resulting in defective hematopoietic support
Blood, May 1, 2004; 103(9): 3349 - 3354.
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