Prospective Isolation and Characterization of Human Bone Marrow-Derived MSCs
There is an increasing interest in adult stem cells, especially mesenchymal stem/stromal cells (MSCs), in hematology and regenerative medicine because of the simplicity of isolation and ex vivo expansion of these cells. Conventionally, MSCs are functional
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rospective Isolation and Characterization of Human Bone Marrow-Derived MSCs A. Harichandan, K. Sivasubramaniyan and H.-J. Bühring Abstract There is an increasing interest in adult stem cells, especially mesenchymal stem/stromal cells (MSCs), in hematology and regenerative medicine because of the simplicity of isolation and ex vivo expansion of these cells. Conventionally, MSCs are functionally isolated from tissue based on their capacity to adhere to the surface of culture flasks. This isolation procedure is hampered by the unpredictable influence of secreted molecules and interactions with co-cultured hematopoietic and other unrelated cells, as well as by the arbitrarily selected removal time of non-adherent cells prior to the expansion of MSCs. Finally, functionally isolated cells do not provide biological information about the starting population. To circumvent these limitations, several strategies have been developed to facilitate the prospective isolation of MSCs based on the selective expression or absence of surface markers. The isolation and ex vivo expansion of these cells require an adequate quality control of the source and product. Here we summarize the most frequently used markers and introduce new targets for antibody-based isolation and characterization of bone marrow-derived MSCs.
Keywords Flow cytometry Mesenchymal/stromal stem cells MSCs Prospective isolation Surface antigens
MSC subsets
Abbreviations MSC SSEA-4 TNAP CFU-F PDGF Rb
Mesenchymal stem/stromal cell Stage specific embryonic antigen 4 Tissue nonspecific alkaline phosphatase Colony forming units—fibroblast Platelet derived growth factor receptor beta
A. Harichandan K. Sivasubramaniyan H.-J. Bühring (&) Division of Haematology, Immunology, Oncology, Rheumatology, and Pulmonology, Department of Internal Medicine II, University Clinic of Tübingen, Tübingen, Germany e-mail: [email protected] H.-J. Bühring Laboratory for Stem Cell Research, Division of Haematology, Oncology, Immunology, and Rheumatology, Department of Internal Medicine II, University Clinic, Otfried-Müller-Str. 10, 72076 Tübingen, Germany
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NK CDCP1 SSEA-3
A. Harichandan et al.
Natural killer cells CUB domain-containing protein 1 Stage specific embryonic antigen 3
Contents 1 2 3
Characteristics of Bone Marrow-Derived MSCs ............................................................... MSCs from Other Tissues................................................................................................... Isolation Procedures of MSCs ............................................................................................ 3.1 Functional Isolation of MSCs .................................................................................... 3.2 Prospective Isolation of MSCs................................................................................... 3.3 Prospective Isolation of MSC Subsets....................................................................... 3.4 Isolation of a Single MSC......................................
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