Protocol

Cellular Programming and Reprogramming

Volume 636 of the series Methods in Molecular Biology pp 55-78

Date:

Isolation Procedure and Characterization of Multipotent Adult Progenitor Cells from Rat Bone Marrow

  • Kartik SubramanianAffiliated withStem Cell Institute and Department of Chemical Engineering and Materials Science, University of Minnesota
  • , Martine GeraertsAffiliated withStem Cell Institute Leuven, Catholic University of Leuven
  • , Karen A. PauwelynAffiliated withStem Cell Institute Leuven, Catholic University of Leuven
  • , Yonsil ParkAffiliated withDepartment of Chemical Engineering and Materials Science and Department of Biomedical Engineering, University of Minnesota
  • , D. Jason OwensAffiliated withStem Cell Institute and Department of Chemical Engineering and Materials Science, University of Minnesota
  • , Manja MuijtjensAffiliated withStem Cell Institute Leuven, Catholic University of Leuven
  • , Fernando Ulloa-MontoyaAffiliated withStem Cell Institute Leuven, Catholic University of LeuvenStem Cell Institute and Department of Chemical Engineering and Materials Science, University of Minnesota
  • , Yeuhua JiangAffiliated withStem Cell Institute Leuven, Catholic University of Leuven
  • , Catherine M. VerfaillieAffiliated withStem Cell Institute Leuven, Catholic University of Leuven Email author 
    • , Wei-Shou HuAffiliated withStem Cell Institute Leuven, Catholic University of Leuven

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Abstract

Multipotent adult progenitor cells (MAPCs) are adult stem cells derived from the bone marrow of mouse and rat and were described for the first time in 2002 (Jiang et al., Nature 418:41-49, 2002), and subsequently (Breyer et al., Exp Hematol 34:1596-1601, 2006; Jiang et al., Exp Hematol 30:896-904, 2002; Ulloa-Montoya et al., Genome Biol 8:R163, 2007). The capacity of rodent MAPC to differentiate at the single-cell level into some of the cell types of endoderm, mesoderm, and neuroectoderm germ layer lineages makes them promising candidates for the study of developmental processes. MAPC are isolated using adherent cell cultures and are selected based on morphology after a period of about 8–18 weeks. Here, we describe a step-by-step reproducible method to isolate rat MAPC from fetal and adult bone marrow. We elaborate on several aspects of the isolation protocol including, cell density and medium components, and methods for selecting and obtaining potential MAPC clones and their characterization.

Key words

Adult stem cells Bone marrow Pluripotency Differentiation