Isolation and Characterization of Multipotent and Pluripotent Stem Cells from Human Peripheral Blood
Ciro Gargiulo1*, Van Hung Pham2, Nguyen Thuy Hai1, Kieu C. D. Nguyen3, Pham Van Phuc4, Kenji Abe5, Veronica Flores6, Melvin Shiffman7
1Division of Internal Medicine, The Human Medicine International Clinic, Ho Chi Minh City, Vietnam.
2Molecular Diagnostics Department, Nam Khoa-Biotek Laboratory, Ho Chi Minh City, Vietnam.
3Division of Pathology, The Human Medicine International Clinic, Ho Chi Minh City, Vietnam.
4Department of Stem Cells and Regenerative Medicine, University of Natural Science, Ho Chi Minh City, Vietnam.
5Department of Pathology, National Institute of Infectious Diseases, Tokyo, Japan.
6Sol Price School of Public Policy, University of Southern California, Los Angeles, USA.
7Section of Surgery, Newport Specialty Hospital, Tustin, USA.
DOI: 10.4236/scd.2015.53003   PDF   HTML   XML   5,937 Downloads   8,345 Views   Citations


Stem cells are commonly classified based on the developmental stage from which they are isolated, although this has been a source of debate amongst stem cell scientists. A common approach classifies stem cells into three different groupings: Embryonic Stem Cells (ESCs), Umbilical Cord Stem Cells (UCBSCs) and Adult Stem Cells (ASCs), which include stem cells from bone marrow (BM), fat tissue (FT), engineered induced pluripotent (IP) and peripheral blood (PB). By definition stem cells are progenitor cells capable of self-renewal and differentiation hypothetically “ab infinitum” into more specialized cells and tissue. The main intent of this study was to determine and characterize the different sub-groups of stem cells present within the human PB-SCs that may represent a valid opportunity in the field of clinical regenerative medicine. Stem cells in the isolated mononucleated cells were characterized using a multidisciplinary approach that was based on morphology, the expression of stem cell markers by flowcytometry and fluorescence analysis, RT-PCR and the capacity to self-renew or proliferate and differentiate into specialized cells. This approach was used to identify the expression of hematopoietic, mesenchymal, embryonic and neural stem cell markers. Both isolated adherent and suspension mononucleated cells were able to maintain their stem cell properties during in-vitro culture by holding their capacity for proliferation and differentiation into osteoblast cells, respectively, when exposed to the appropriate induction medium.

Share and Cite:

Gargiulo, C. , Pham, V. , Thuy Hai, N. , Nguyen, K. , Phuc, P. , Abe, K. , Flores, V. and Shiffman, M. (2015) Isolation and Characterization of Multipotent and Pluripotent Stem Cells from Human Peripheral Blood. Stem Cell Discovery, 5, 19-32. doi: 10.4236/scd.2015.53003.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Ab Kadir, R., Hisham Zainal Ariffin, S., Megat Abdul Wahab, R. and Senafi, S. (2012) Molecular Characterisation of Human Peripheral Blood Stem Cells. South African Journal of Science, 108, 1-7.
[2] Trivanovic, D., Kocic, J., Mojsilovic, S., Krstic, A., Ilic, V., Djordjevic, I.O., Santibanez, J.F., et al. (2013) Mesenchymal Stem Cells from Peripheral Blood and Umbilical Cord Wharton’s Jelly. Srpski arhiv za celokupno lekarstvo, 141, 178-186.
[3] Bianco, P., Riminucci, M., Gronthos, S. and Robey, P.G. (2001) Bone Marrow Stromalcells: Nature, Biology, and Potential Applications. Stem Cells, 19, 180-192.
[4] Caplan, A.I. (2007) Adult Mesenchymal Stem Cells for Tissue Engineering versus Regenerative Medicine. Journal of Cellular Physiology, 213, 341-347.
[5] Chamberlain, G., Fox, J., Ashton, B. and Middleton, J. (2007) Mesenchymal Stem Cells: Their Phenotype, Differentiation Capacity, Immunological Features, and Potential for Homing. Stem Cells, 25, 2739-2749.
[6] Gimble, J.M., Guilak, F., Nuttall, M.E., Sathishkumar, S., Vidal, M. and Bunnell, B.A. (2008) In Vitro Differentiation Potential of Mesenchymal Stem Cells. Transfusion Medicine and Hemotherapy, 35, 228-238.
[7] Wagner, W., Wein, F., Seckinger, A., Frankhauser, M., Wirkner, U., Krause, U., et al. (2005) Comparative Characteristics of Mesenchymal Stem Cells from Human Bone Marrow, Adipose Tissue, and Umbilical Cord Blood. Experimental Hematology, 33, 1402-1416.
[8] Kassis, I., Zangi, L., Rivkin, R., Levdansky, L., Samuel, S., Marx, G., et al. (2006) Isolation of Mesenchymal Stem Cells from G-CSF-Mobilized Human Peripheral Blood Using Fibrin Microbeads. Bone Marrow Transplant, 37, 967-976.
[9] Sakaguchi, Y., Sekiya, I., Yaqishita, K. and Muneta, T. (2005) Comparison of human Stem Cells Derived from Various Mesenchymal Tissues. Arthritis & Rheumatism, 52, 2521-2529.
[10] Gronthos, S., Mankani, M., Brahim, J., Robey, P.G. and Shi, S. (2000) Postnatal Human Dental Pulp Stem Cells (DPSCs) in Vitro and in Vivo. Proceedings of the National Academy of Sciences of the United States of America, 97, 25-30.
[11] Wang, H.S., Hung, S.C., Peng, S.T., Huang, C.C., Wei, H.M., Guo, Y.J., et al. (2004) Mesenchymal Stem Cells in the Wharton’s Jelly Umbilical Cord. Stem Cells, 22, 1330-1337.
[12] Locke, M., Feisst, V. and Dunbar, P.R. (2011) Concise Review. Human Adipose Derived Stem Cells: Separating Promise from Clinical Need. Stem Cells, 29, 404-411.
[13] Patel, A.N. and Genovese, J. (2011) Potential Clinical Applications of Adult Human Mesenchymal Stem Cell (Prochymal®) Therapy. Stem Cells and Cloning: Advances and Applications, 4, 61-72.
[14] Domen, J., Wagers, A. and Weissman, I.L. (2006) Regenerative Medicine Bone Marrow (Hematopoietic) Stem Cells. National Institute of Health, 2, 14-28.
[15] Sun, Y., Kong, W., Falk, A., Hu, J., Zhou, L., Pollard, S. et al. (2009) CD133 (Prominin) Negative Human Neural Stem Cells Are Clonogenic and Tripotent. PLoS ONE, 4, e5498.
[16] Callas, A., Pook, M., Treika, A. and Maimets, T. (2014) SOX2 Is Regulated Differently from NANOG and OCT4 in Human Embryonic Stem Cells during Early Differentiation Initiated with Sodium Butyrate. Stem Cells International, 2014, Article ID: 298163.
[17] Chong, P.P., Selvaratnam, L., Abbas, A.A. and Kamarul, T. (2011) Human Peripheral Blood Derived Mesenchymal Stem Cellsdemonstrate Similar Characteristics and Chondrogenic Differentiation Potential to Bone Marrow Derived Mesenchymal Stem Cells. Journal of Orthopedic Research, 30, 634-642.
[18] Aubin, J.E. and Heersche, J.N. (2000) Osteoprogenitor Cell Differentiation to Mature Bone Forming Osteoblasts. Drug Development Research, 49, 206-215.<206::AID-DDR11>3.0.CO;2-G
[19] Undale, A.H., Westerndorf, J.J., Yaszemski, M.J. and Khosla, S. (2009) Mesenchymal Stem Cells for Bone Repair and Metabolic Bone Disease. Mayo Clinic Proceedings, 84, 893-902.
[20] Aubin, J.E., Malaval, F. and Gupta, A.K. (1995) Osteoblasts and Chondroblasts Differentiation. Bone, 17, 77-83.
[21] Homicz, M.R., Schumacher, B.L., Sah, R.L. and Watson, D. (2012) Effects of Serial Expansion of Septal Chondrocytes on Tissue-Engineered Neocartilage Composition. Otolaryngology Head and Neck Surgery, 127, 398-408.
[22] Gang, H., Peng, L., Jie, F. and Yan, J. (2011) A Novel Population of Mesenchymal Progenitors with Hematopoietic Potential Originated from CD14‾ Peripheral Blood Mononuclear Cells. International Journal of Medical Sciences, 8, 16-29.
[23] Grage-Griebenow, E., Flad, H.D. and Ernst, M. (2001) Heterogeneity of Human Peripheral Blood Monocyte Subsets. Journal of Leukocyte Biology, 69, 11-20.
[24] Lane, L.W., Williams, D.A. and Watt, F.M. (2014) Mod-ulating the Stem Cell Niche for Tissue Regeneration. Nature Biotechnology, 32, 795-803.
[25] Havens, A.M., Shiozawa, Y., Jung, Y., Sun, H., Wang, J., McGee, S., et al. (2012) Human Very Small Embryonic-Like Cellsgenerate Skeletal Structures, in Vivo. Stem Cells and Development, 1, 1-9.
[26] Haider, H.K., Jiang, S., Idris, N.M. and Ashraf, M. (2008) IGF-1-Overexpressing Mesenchymal Stem Cells Accelerate Bone Marrow Stem Cell Mobilization via Paracrine Activation of SDF-1α/CXCR4 Signaling to Promote Myocardial Repair. Circulation Research, 103, 1300-1308.
[27] Senthil, P. (2013) Adult Stem Cell and Embryonic Stem Cell Markers. Mater Methods, 3, 200.
[28] Zhao, W., Ji, X., Zhang, F., Li, L. and Ma, L. (2012) Embryonic Stem Cell Markers. Molecules, 17, 6196-6236.
[29] Draper, J.S., Pigott, C., Thomson, J.A. and Andrews, P.W. (2002) Surface Antigens of Human Embryonic Stem Cells: Changes upon Differentiation in Culture. Journal of Anatomy, 200, 249-258.
[30] Xu, C.H., Inokuma, M.S., Denham, J., Golds, K., Kundu, P., Gold, J.D., et al. (2001) Feeder-Free Growth of Undifferentiated Human Embryonic Stem Cells. Natural Biotechnology, 19, 971-974.
[31] Skottman, H., Mikkola, M., Lundin, K., Olsson, C., Strömberg, A.M., Tuuri, T., et al. (2005) Gene Expression Signatures of Seven Individual Human Embryonic Stem Cell Lines. Stem Cells, 23, 1343-1356.
[32] Mingyu, Z., Tao, S., Liang, Y., Chen, R., Wu, L., Yang, Z., et al. (2008) Nestin and CD133; Valuable Stem Cell-Specific Markers for Determining Clinical Outcome of Glioma Patients. Journal of Experimental & Clinical Cancer Research, 27, 85-92.
[33] Knoepfler, P. (2009) Deconstructing Stem Cell Tumorigenicity: A Roadmap to Safe Regenerative Medicine. Stem Cells, 27, 1050-1056.
[34] Schurch, C.M., Riether, C. and Ochsenbein, A.F. (2014) Cytotoxic CD8+ T Cells Stimulate Hematopoietic Progenitors by Promoting Cytokine Release from Bone Marrow Mesenchymal Stromal Cells. Cell Press, 14, 460-472.
[35] Weiner, R.S. and Kincade, P.W. (2014) 9-1-1: HSCs Respond to Emergency Calls. Cell Press, 14, 415-416.
[36] Zhong, Z.N., Zhu, S.F., Yuan, A.D., Lu, G.H., He, Z.Y., Fa, Z.Q., et al. (2012) Potential of Placenta-Derived Mesenchymal Stem Cells as Seed Cells for Bone Tissue Engineering: Preliminary Study of Osteoblastic Differentiation and Immunogenicity. Orthopedics, 35, 779-788.
[37] Rossi, L., Salvestrini, V., Ferrari, D., Virgilio, F.D., Lemoli, R.M., et al. (2012) The Sixth Sense: Hematopoietic Stem Cells Detect Danger through Purinergic Signaling. Blood, 120, 2365-2375.
[38] Rosland, G.V., Svendsen, A., Torsvik, A., Sobala, E., McCormack, E., Immervoll, H., et al. (2009) Long-Term Cultures of Bone Marrow-Derived Human Mesenchymal Stem Cells Frequently Undergo Spontaneous Malignant Transformation. Cancer Research, 69, 5331-5339.
[39] Izadpanah, R., Kaushal, D., Kriedt, C., Tsien, F., Patel, B., Dufour, J., et al. (2008) Long-Term in Vitro Expansion Alters the Biology of Adult Mesenchymal Stem Cells. Cancer Research, 68, 4229-4238.

Copyright © 2023 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.