Elsevier

Tissue and Cell

Volume 41, Issue 6, December 2009, Pages 448-455
Tissue and Cell

Short communication
Characterization of mesenchymal stem cells (MSCs) from human fetal lung: Potential differentiation of germ cells

https://doi.org/10.1016/j.tice.2009.05.004Get rights and content

Abstract

Pluripotent mesenchymal stem-like cell lines were established from lungs of 3–4 months old aborted fetus. The cells present the high ex vivo expansion potential of MSC, a typical fibroblast-like morphology and proliferate up to 15 passages without displaying clear changes in morphology. Immunological localization and flow cytometry analyses showed that these cells are positive for OCT4, c-Kit, CD11, CD29, CD44, telomerase, CD106, CD105, CD166, and SSEA1, weakly expression or negative for SSEA1, SSEA3, SSEA4, CD34, CD105 and CD106. These cells can give rise to the adipogenic as evidenced by accumulation of lipid-rich vacuoles within cells identified by Oil-red O when they were induced with 0.5 mM isobutylmethylxanthine, 200 μM indomethacin, 10−6 M dexamethasone, and 10 μg/ml of insulin in high-glucose DMEM. Osteogenic lineage cells were generated in 0.1 μM dexamethasone, 50 μg/ml ascorbic acid, 10 mM β-glycerophosphate, which are shaped as the osteoblastic morphology, expression of alkaline phosphatase (AP), and the formation of a mineralized extracellular matrix identified by Alizarin Red staining. Neural cells are observed when the cultures were induced with 2-mercapometal, which are positive for nestin, NF-100, MBP and GFAP. Additionally, embryoid bodies (EBs) and sperm like cells are obtained in vitro differentiation of these lung MSCs induced with 10−5 M retinoic acid (RA). These results demonstrated that these MSCs are pluripotent and may provide an in vitro model to study germ-cell formation and also as a potential source of sperms for male infertility.

Introduction

Mesenchymal stem cells (MSCs) are multipotent progenitor cells with the capacity to differentiate into many types of tissue cells (Pittenger et al., 1999, Hennrick et al., 2007, Young et al., 2001). Bone marrow is believed to be the enriched reservoir of MSCs and major source for these precursor cells that populate other adult tissues and organs (Prockop, 1997, Majumdar et al., 1998). MSCs can be isolated from circulating blood (Zvaifler et al., 2000), as well as from diverse nonhematopoietic tissues and connective tissues such as synovium (DeBari et al., 2001), adipose tissue (Zuk et al., 2001), trabecular bone (Noth et al., 2002), dermis (Young et al., 2001, Hoogduijn et al., 2007), dental pulp (Pierdomenico et al., 2005), and the lung (Sabatini et al., 2005). There have been no reports of MSCs that have been conclusively demonstrated to be of tissue-specific origin in any adult organ system, including the lung (Lama et al., 2007). The origins of mesenchymal cells that participate in normal and abnormal repair responses in the human adult lung remain unclear. In this study, we isolate the multipotent MSCs from the human fetal lung and investigate the ability to differentiate into dipocytes and osteoblasts, even cardiac cells, neural cells and sperm like cells.

Section snippets

Samples

The study was approved by the patient and the biological ethical committee of Shaanxi Centre of Stem Cells Engineering & Technology. The lung were mechanically collected, washed with PBS again, dissected and incubated with 0.125% trypsin and 0.02% EDTA for 10–20 min or with 0.125% collagenase IV for 20–40 min at 37 °C, then gently pipetted, centrifuged at 1000 rpm for 5–10 min. Cells were seeded at a density of 1 × 105 mononuclear cells per microliter and transferred to medium containing 10% FCS

Characterization of hMSCs derived human fetal lung

Isolation of plastic-adherent fibroblastoid cells from human fetal lung. MSCs have been well recognized to derive from single-cell suspensions of bone marrow by the selective growth of plastic-adherent fibroblast-like cell colonies in liquid culture medium. Within 2–5 passages after the initial plating of the primary culture, hMSCs derived human fetal lung appeared as a monolayer of large, flat, and spindle-shaped cells (Fig. 1A). As the cells approached confluence, they assumed a more

Discussion

In this study, we investigated the possibility to isolate fibroblast-like cells from human fetal lung and these cells exhibited several characteristics typical of classical bone marrow derived MSCs (Pittenger et al., 1999, Short et al., 2003, Majumdar et al., 1998). Firstly, they can be isolated by adherence purification on tissue culture plastic and form distinct CFU-Fs in vitro culture; secondly, these cells express surface proteins typically associated with MSCs but do not express

Acknowledgments

We are grateful to Professor Huayan Wang for discussion on the manuscript. This work was supported by grants from the Program (30200137) from National Natural Science Foundation of China, Key Program of State Education Ministry (109148), The Scientific Research Program of Shaanxi Province (2008K02-05), China Postdoctoral Science Foundation funded project (20080431253, 200801438), the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry (14110101)

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