Isolation Procedure and Characterization of Multipotent Adult Progenitor Cells from Rat Bone Marrow
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 wordsAdult stem cells Bone marrow Pluripotency Differentiation
The authors would like to thank Dr Hirsch and LeAnn Oseth of the University of Minnesota cytogenetics core facility for accurate performance and analysis of G-banding experiments. Tine Decuyper, Sarah Mertens, Nathalie Feyaerts, Annelies Deman, April Breyer, Tineke Notelaers, Pieter Berckmans, and Sumitha Jacob are acknowledged for their excellent technical support with cell culture, stainings and qRT-PCR. Vick Vanduppen is acknowledged for his FACS support. This work was supported by grants to CMV: NIH-T15 HL076653, KUL CoE, FWO Odysseus Fund; FP6-STREP: STROKEMAP. KS was funded by Doctoral dissertation fellowship (DDF), Graduate School, University of Minnesota. MG was funded by FWO; DJO was supported by the NIH biotechnology Training Grant (GM08347); KP was funded by the FWO, BAEF, and VVGE.
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