Wexler Sarah A, Donaldson Craig, Denning-Kendall Patricia, Rice Claire, Bradley Ben, Hows Jill M
Department of Transplantation Sciences, Paul O'Gorman Lifeline Centre, Division of Medicine, University of Bristol, Bristol, UK.
Br J Haematol. 2003 Apr;121(2):368-74. doi: 10.1046/j.1365-2141.2003.04284.x.
In postnatal life, mesenchymal stem cells (MSC) self-replicate, proliferate and differentiate into mesenchymal tissues, including bone, fat, tendon, muscle and bone marrow (BM) stroma. Possible clinical applications for MSC in stem cell transplantation have been proposed. We have evaluated the frequency, phenotype and differentiation potential of MSC in adult BM, cord blood (CB) and peripheral blood stem cell collections (PBSC). During culture, BM MSC proliferated to confluence in 10-14 d, maintaining a stable non-haemopoietic phenotype, HLA class-1+, CD29+, CD44+, CD90+, CD45-, CD34- and CD14 through subsequent passages. Using the colony forming unit fibroblasts assay, the estimated frequency of MSC in the BM nucleated cell population was 1 in 3.4 x 10(4) cells. Both adipogenic and osteogenic differentiation of BM MSC was demonstrated. In contrast, CB and PBSC mononuclear cells cultured in MSC conditions for two passages produced a population of adherent, non-confluent fibroblast-like cells with a haemopoietic phenotype, CD45+, CD14+, CD34-, CD44-, CD90- and CD29-. In paired experiments, cultured BM MSC and mature BM stroma were seeded with CB cells enriched for CD34+. Similar numbers of colony-forming units of granulocytes-macrophages were produced by MSC-based and standard stroma cultures over 10 weeks. We conclude that adult BM is a reliable source of functional cultured MSC, but CB and PBSC are not.
在出生后的生命中,间充质干细胞(MSC)自我复制、增殖并分化为间充质组织,包括骨骼、脂肪、肌腱、肌肉和骨髓(BM)基质。已经有人提出了MSC在干细胞移植中的潜在临床应用。我们评估了成人BM、脐血(CB)和外周血干细胞采集物(PBSC)中MSC的频率、表型和分化潜能。在培养过程中,BM MSC在10 - 14天内增殖至汇合,在随后的传代过程中维持稳定的非造血表型,HLA - 1+、CD29+、CD44+、CD90+、CD45-、CD34-和CD14-。使用集落形成单位成纤维细胞试验,估计BM有核细胞群体中MSC的频率为3.4×10⁴个细胞中有1个。BM MSC的成脂和成骨分化均得到证实。相比之下,在MSC条件下培养两代的CB和PBSC单核细胞产生了一群具有造血表型CD45+、CD14+、CD34-、CD44-、CD90-和CD29-的贴壁、未汇合的成纤维细胞样细胞。在配对实验中,用富集CD34+的CB细胞接种培养的BM MSC和成熟BM基质。在10周内,基于MSC的培养和标准基质培养产生的粒细胞 - 巨噬细胞集落形成单位数量相似。我们得出结论,成人BM是功能性培养MSC的可靠来源,但CB和PBSC不是。