从大鼠骨髓中分离多能成体祖细胞的方法及特性研究

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

作者信息

Subramanian Kartik, Geraerts Martine, Pauwelyn Karen A, Park Yonsil, Owens D Jason, Muijtjens Manja, Ulloa-Montoya Fernando, Jiang Yeuhua, Verfaillie Catherine M, Hu Wei-Shou

机构信息

Department of Chemical Engineering and Materials Science, Stem Cell Institute, University of Minnesota, Minneapolis, MN, USA.

出版信息

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

DOI:10.1007/978-1-60761-691-7_4

PMID:20336516

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.

摘要

多能成体祖细胞（MAPCs）是源自小鼠和大鼠骨髓的成体干细胞，于2002年首次被描述（Jiang等人，《自然》418:41 - 49，2002年），随后也有相关报道（Breyer等人，《实验血液学》34:1596 - 1601，2006年；Jiang等人，《实验血液学》30:896 - 904，2002年；Ulloa - Montoya等人，《基因组生物学》8:R163，2007年）。啮齿动物MAPC在单细胞水平上分化为内胚层、中胚层和神经外胚层胚层谱系的某些细胞类型的能力，使其成为发育过程研究中有前景的候选细胞。MAPC通过贴壁细胞培养进行分离，并在约8 - 18周的一段时间后根据形态进行筛选。在此，我们描述一种从胎儿和成年骨髓中分离大鼠MAPC的逐步可重复方法。我们详细阐述了分离方案的几个方面，包括细胞密度和培养基成分，以及选择和获得潜在MAPC克隆及其鉴定的方法。