A novel rich source of human mesenchymal stem cells from the debris of bone marrow samples

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Abstract

The debris from human bone marrow (BM) samples is generally filtered out and discarded prior to isolation of mesenchymal stem cells (MSCs). The purpose of this study is to develop a method to harvest MSCs from the debris and investigate their biological characteristics compared with the marrow counterparts. The BM tissue fragments were digested with collagenase and this treatment yielded mononuclear cells half to those from the corresponding filtered BM. The frequencies of colony-forming unit-fibroblast in these two cell populations were not significantly different. MSCs of two origins exhibited similar morphological and phenotypic features. Fluorescent dye-dilution assay showed that they grew at comparable rates both in the primary and passaging cultures. Further, they could be induced into osteoblasts, chondroblasts and adipocytes, as revealed by histological and molecular examinations. Thus, BM tissue fragments may serve as a new source of MSCs in the settings of bench experiments and clinical trials.

Section snippets

Materials and methods

Isolation and culture of MSC form BM debris and BM. BM was harvested from 7 donors (three women and four men at the age ranging from 28 to 58 years,) following informed consent according to the guideline from Beijing Institute of Radiation Medicine Committee on the Use of Human Subjects in Research. Collected marrow was passed through a cell-strainer with the pore size of 80 μm in diameter (BD, USA). Mononuclear cells were isolated from the filtered BM by the routine density gradient

Results

To observe if MSCs existed in the debris of human BM samples, the tissue fragments were digested with collagenase and mononuclear cells were harvested by gradient centrifugation. The proportion of mononuclear cells was about 80% in the cell suspension from the digested BM debris, which was obviously higher than that in the BM analogue. Moreover, typically, the number of mononuclear cells in the digested debris could reach up to half of that from the filtered marrow.

When the digested cells were

Discussion

Although MSCs can be isolated from many tissues, bone marrow is a reproducible and convenient source of these cells from nearly all species tested except mice [11]. In this study, we tried to isolate MSCs from the tissue fragments in the human marrow samples, which are mainly composed of adipose tissue, compact bone fragments and vessel networks. Our results here suggest that these cells meet the generally accepted standard criteria including the fibroblast-like morphology, the expression of

Acknowledgments

This research was supported by Chinese High-tech Grant (No. 2007AA02Z454) and Beijing Medical Development Grants (No. 2007-2033 and No. 2006-2042).

References (21)

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