Isolation procedure and characterization of multipotent adult progenitor cells from rat bone marrow

Methods Mol Biol. 2010:636:55-78. doi: 10.1007/978-1-60761-691-7_4.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult Stem Cells* / cytology
  • Adult Stem Cells* / physiology
  • Animals
  • Biomarkers / metabolism
  • Bone Marrow Cells* / cytology
  • Bone Marrow Cells* / physiology
  • Cell Culture Techniques*
  • Cell Differentiation
  • Cell Line
  • Cell Separation / instrumentation
  • Cell Separation / methods*
  • Female
  • Flow Cytometry / methods
  • Gene Expression Profiling
  • Humans
  • Mice
  • Multipotent Stem Cells* / cytology
  • Multipotent Stem Cells* / physiology
  • Pregnancy
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Inbred F344
  • Rats, Sprague-Dawley

Substances

  • Biomarkers
  • RNA, Messenger