Abstract
Bone marrow-derived cells have been postulated as a source of multipotent mesenchymal stem cells (MSC). However, the whole fraction of MSC remains heterogeneous and the expansion of primitive subset of these cells is still not well established. Here, we optimized the protocol for propagating the low-adherent subfraction of MSC which results in long-term expansion of population characterized by CD45−CD14+CD34+ phenotype along with expression of common MSC markers. We established that the expanded MSC are capable of differentiating into endothelial cells highly expressing angiogenic markers and exhibiting functional properties of endothelium. Moreover, we found these cells to be multipotent and capable of giving rise into cells from neuronal lineages. Interestingly, the expanded MSC form characteristic cellular spheres in vitro indicating primitive features of these cells. In sum, we isolated the novel multipotent subpopulation of CD45−CD14+ CD34+ bone marrow-derived cells that could be maintained in long-term culture without losing this potential.
Similar content being viewed by others
References
Abdel-Latif, A., Bolli, R., Tleyjeh, I.M., Montori, V.M., Perin, E.C., Hornung, C.A., Zuba-Surma, E.K., Al-Mallah, M., and Dawn, B. (2007). Adult bone marrow-derived cells for cardiac repair: a systematic review and meta-analysis. Arch. Intern. Med. 167, 989–997.
Alviano, F., Fossati, V., Marchionni, C., Arpinati, M., Bonsi, L., Franchina, M., Lanzoni, G., Cantoni, S., Cavallini, C., Bianchi, F., et al. (2007). Term Amniotic membrane is a high throughput source for multipotent Mesenchymal Stem Cells with the ability to differentiate into endothelial cells in vitro. BMC Dev. Biol. 7, 11.
Beyer Nardi, N., and da Silva Meirelles, L., (2006). Mesenchymal stem cells: isolation, in vitro expansion and characterization. Handb. Exp. Pharmacol. 174, 249–282.
Brunt, K.R., Hall, S.R.R., Ward, C.A., and Melo, L.G. (2007). Endothelial progenitor cell and mesenchymal stem cell isolation, characterization, viral transduction. Methods Mol. Med. 139, 197–210.
Chawla, A., Schwarz, E.J., Dimaculangan, D.D., and Lazar, M.A. (1994). Peroxisome proliferator-activated receptor (PPAR) gamma: adipose-predominant expression and induction early in adipocyte differentiation. Endocrinology 135, 798–800.
Chen, M., Lie, P., Li, Z., and Wei, X. (2009). Endothelial differentiation of Wharton’s jelly-derived mesenchymal stem cells in comparison with bone marrow-derived mesenchymal stem cells. Exp. Hematol. 37, 629–640.
Cleaver, O., and Melton, D.A. (2003). Endothelial signaling during development. Nat. Med. 9, 661–668.
Copland, I., Sharma, K., Lejeune, L., Eliopoulos, N., Stewart, D., Liu, P., Lachapelle, K., and Galipeau, J. (2008). CD34 expression on murine marrow-derived mesenchymal stromal cells: impact on neovascularization. Exp. Hematol. 36, 93–103.
Dawn, B., Tiwari, S., Kucia, M.J., Zuba-Surma, E.K., Guo, Y., Sanganalmath, S.K., Abdel-Latif, A., Hunt, G., Vincent, R.J., Taher, H., et al. (2008). Transplantation of bone marrow-derived very small embryonic-like stem cells attenuates left ventricular dysfunction and remodeling after myocardial infarction. Stem Cells 26, 1646–1655.
Dawn, B., Abdel-Latif, A., Sanganalmath, S.K., Flaherty, M.P., Zuba-Surma, E.K. (2009). Cardiac repair with adult bone marrowderived cells: the clinical evidence. Antioxid. Redox Signal. 11, 1865–1882.
D’Ippolito, G., Diabira, S., Howard, G.A., Menei, P., Roos, B.A., and Schiller, P.C. (2004). 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. 117, 2971–2981.
Fei, R.G., Penn, P.E., and Wolf, N.S. (1990). A method to establish pure fibroblast and endothelial cell colony cultures from murine bone marrow. Exp. Hematol. 18, 953–957.
Fernandez Pujol, B., Lucibello, F.C., Gehling, U.M., Lindemann, K., Weidner, N., Zuzarte, M.L., Adamkiewicz, J., Elsässer, H.P., Müller, R., and Havemann, K. (2000). Endothelial-like cells derived from human CD14 positive monocytes. Differentiation 65, 287–300.
Gang, E.J., Jeong, J.A., Han, S., Yan, Q., Jeon, C., and Kim, H. (2006). In vitro endothelial potential of human UC blood-derived mesenchymal stem cells. Cytotherapy 8, 215–227.
Girdlestone, J., Limbani, V., Cutler, A., and Navarrete, C. (2009). Efficient expansion of mesenchymal stromal cells from umbilical cord under low serum conditions. Cytotherapy [Epub ahead of print].
Gnecchi, M., and Melo, L.G. (2009). Bone marrow-derived mesenchymal stem cells: isolation, expansion, characterization, viral transduction, and production of conditioned medium. Methods Mol. Biol. 482, 281–294.
Grove, J.E., Bruscia, E., and Krause, D.S. (2004). Plasticity of bone marrow-derived stem cells. Stem Cells 22, 487–500.
Hofstetter, C.P., Schwarz, E.J., Hess, D., Widenfalk, J., El Manira, A., Prockop, D.J., and Olson, L. (2002). Marrow stromal cells form guiding strands in the injured spinal cord and promote recovery. Proc. Natl. Acad. Sci. USA 99, 2199–2204.
Houlihan, D.D., and Newsome, P.N. (2008). Critical review of clinical trials of bone marrow stem cells in liver disease. Gastroenterology 135, 438–450.
Ingram, D.A., Mead, L.E., Tanaka, H., Meade, V., Fenoglio, A., Mortell, K., Pollok, K., Ferkowicz, M.J., Gilley, D., and Yoder, M.C. (2004). Identification of a novel hierarchy of endothelial progenitor cells using human peripheral and umbilical cord blood. Blood 104, 2752–2760.
Jain, R.K. (2003). Molecular regulation of vessel maturation. Nat. Med. 9, 685–693.
Jiang, Y., Jahagirdar, B.N., Reinhardt, R.L., Schwartz, R.E., Keene, C.D., Ortiz-Gonzalez, X.R., Reyes, M., Lenvik, T., Lund, T., Blackstad, M., et al. (2002). Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 418, 41–49.
Kabos, P., Ehtesham, M., Kabosova, A., Black, K.L., and Yu, J.S. (2002). Generation of neural progenitor cells from whole adult bone marrow. Exp. Neurol. 178, 288–293.
Kaiser, S., Hackanson, B., Follo, M., Mehlhorn, A., Geiger, K., Ihorst, G., and Kapp, U. (2007). BM cells giving rise to MSC in culture have a heterogeneous CD34 and CD45 phenotype. Cytotherapy 9, 439–450.
Karussis, D., Kassis, I., Kurkalli, B.G.S., and Slavin, S. (2008). Immunomodulation and neuroprotection with mesenchymal bone marrow stem cells (MSCs): a proposed treatment for multiple sclerosis and other neuroimmunological/neurodegenerative diseases. J. Neurol. Sci. 265, 131–135.
Kopher, R.A., Penchev, V.R., Islam, M.S., Hill, K.L., Khosla, S., and Kaufman, D.S. (2010). Human embryonic stem cell-derived CD34+ cells function as MSC progenitor cells. Bone 47, 718–728.
Kucia, M., Reca, R., Campbell, F.R., Zuba-Surma, E., Majka, M., Ratajczak, J., and Ratajczak, M.Z. (2006). A population of very small embryonic-like (VSEL) CXCR4(+)SSEA-1(+)Oct-4+ stem cells identified in adult bone marrow. Leukemia 20, 857–869.
Kucia, M., Wysoczynski, M., Ratajczak, J., and Ratajczak, M.Z. (2008). Identification of very small embryonic like (VSEL) stem cells in bone marrow. Cell Tissue Res. 331, 125–134.
Kuwana, M., Okazaki, Y., Kodama, H., Izumi, K., Yasuoka, H., Ogawa, Y., Kawakami, Y., and Ikeda, Y. (2003). Human circulating CD14+ monocytes as a source of progenitors that exhibit mesenchymal cell differentiation. J. Leukoc. Biol. 74, 833–845.
Li, Y., Zhang, C., Xiong, F., Yu, M., Peng, F., Shang, Y., Zhao, C., Xu, Y., Liu, Z., Zhou, C., et al. (2008). Comparative study of mesenchymal stem cells from C57BL/10 and mdx mice. BMC Cell Biol. 9, 24.
Lim, S.Y., Kim, Y.S., Ahn, Y., Jeong, M.H., Hong, M.H., Joo, S.Y., Nam, K.I., Cho, J.G., Kang, P.M., and Park, J.C. (2006). The effects of mesenchymal stem cells transduced with Akt in a porcine myocardial infarction model. Cardiovasc. Res. 70, 530–542.
Martin-Rendon, E., Brunskill, S.J., Hyde, C.J., Stanworth, S.J., Mathur, A., and Watt, S.M. (2008). Autologous bone marrow stem cells to treat acute myocardial infarction: a systematic review. Eur. Heart J. 29, 1807–1818.
Matoba, S., Tatsumi, T., Murohara, T., Imaizumi, T., Katsuda, Y., Ito, M., Saito, Y., Uemura, S., Suzuki, H., Fukumoto, S., et al. (2008). Long-term clinical outcome after intramuscular implantation of bone marrow mononuclear cells (therapeutic angiogenesis by cell transplantation [TACT] trial) in patients with chronic limb ischemia. Am. Heart J. 156, 1010–1018.
Oswald, J., Boxberger, S., Jørgensen, B., Feldmann, S., Ehninger, G., Bornhäuser, M., and Werner, C. (2004). Mesenchymal stem cells can be differentiated into endothelial cells in vitro. Stem Cells 22, 377–384.
Phinney, D.G., and Prockop, D.J. (2007). Concise review: mesenchymal stem/multipotent stromal cells: the state of transdifferentiation and modes of tissue repair-current views. Stem Cells 25, 2896–2902.
Pitas, R.E., Boyles, J., Mahley, R.W., and Bissell, D.M. (1985). Uptake of chemically modified low density lipoproteins in vivo is mediated by specific endothelial cells. J. Cell Biol. 100, 103–117.
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. (1999). Multilineage potential of adult human mesenchymal stem cells. Science 284, 143–147.
Quirici, N., Soligo, D., Caneva, L., Servida, F., Bossolasco, P., and Deliliers, G.L. (2001). Differentiation and expansion of endothelial cells from human bone marrow CD133(+) cells. Br. J. Haematol. 115, 186–94.
Reynolds, B.A., and Weiss, S. (1992). Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255, 1707–1710.
Romagnani, P., Annunziato, F., Liotta, F., Lazzeri, E., Mazzinghi, B., Frosali, F., Cosmi, L., Maggi, L., Lasagni, L., Scheffold, A., et al. (2005). CD14+CD34 low cells with stem cell phenotypic and functional features are the major source of circulating endothelial progenitors. Circ. Res. 97, 314–322.
Shiota, M., Heike, T., Haruyama, M., Baba, S., Tsuchiya, A., Fujino, H., Kobayashi, H., Kato, T., Umeda, K., Yoshimoto, M., et al. (2007). Isolation and characterization of bone marrow-derived mesenchymal progenitor cells with myogenic and neuronal properties. Exp. Cell Res. 313, 1008–1023.
Torrente, Y., and Polli, E. (2008). Mesenchymal stem cell transplantation for neurodegenerative diseases. Cell Transplant 17, 1103–1113.
Wang, Q.R., Wang, B.H., Huang, Y.H., Dai, G., Li, W.M., and Yan, Q. (2008). Purification and growth of endothelial progenitor cells from murine bone marrow mononuclear cells. J. Cell Biochem. 103, 21–29.
Zhang, S.J., Zhang, H., Hou, M., Zheng, Z., Zhou, J., Su, W., Wei, Y., and Hu, S. (2007). Is it possible to obtain “true endothelial progenitor cells” by in vitro culture of bone marrow mononuclear cells? Stem Cells Dev. 16, 683–690.
Author information
Authors and Affiliations
Corresponding author
About this article
Cite this article
Szade, K., Zuba-Surma, E., Rutkowski, A.J. et al. CD45−CD14+CD34+ murine bone marrow low-adherent mesenchymal primitive cells preserve multilineage differentiation potential in long-term in vitro culture. Mol Cells 31, 497–507 (2011). https://doi.org/10.1007/s10059-011-2176-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10059-011-2176-y