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Mouse Embryonic Fibroblasts (MEF) Exhibit a Similar but not Identical Phenotype to Bone Marrow Stromal Stem Cells (BMSC)

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Abstract

Mouse embryonic fibroblasts have been utilized as a surrogate stem cell model for the postnatal bone marrow-derived stromal stem cells (BMSC) to study mesoderm-type cell differentiation e.g. osteoblasts, adipocytes and chondrocytes. However, no formal characterization of MEF phenotype has been reported. Utilizing standard in vitro and in vivo assays we performed a side-by-side comparison of MEF and BMSC to determine their ability to differentiate into mesoderm-type cells. BMSC were isolated from 8–10 weeks old mouse bone marrow by plastic adherence. MEF were established by trypsin/EDTA digestion from E13.5 embryos after removing heads and viscera, followed by plastic adherence. Compared to BMSC, MEF exhibited telomerase activity and improved cell proliferation as assessed by q-PCR based TRAP assay and cell number quantification, respectively. FACS analysis revealed that MEF exhibited surface markers characteristic of the BMSC: Sca-1+, CD73+, CD105+, CD29+, CD44+, CD106+, CD11b−, and CD45−. In contrast to BMSC, ex vivo osteoblast (OB) differentiation of MEF exhibited a less mature osteoblastic phenotype (less alkaline phosphatase, collagen type I and osteocalcin) as assessed by real-time PCR analysis. Compared to BMSC, MEF exhibited a more enhanced differentiation into adipocyte and chondrocyte lineages. Interestingly, both MEF and BMSC formed the same amount of heterotopic bone and bone marrow elements upon in vivo subcutaneous implantation with hydroxyapatite/tricalcium phosphate, in immune deficient mice. In conclusion, MEF contain a population of stem cells that behave in ex vivo and in vivo assays, similar but not identical, to BMSC. Due to their enhanced cell growth, they may represent a good alternative for BMSC in studying molecular mechanisms of stem cell commitment and differentiation to osteoblasts, adipocytes and chondrocytes.

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Acknowledgements

The authors are grateful to Lone Christiansen for excellent technical assistance. The study was supported by grants from the Lundbeck foundation, the NovoNordisk Foundation, the Karen Elise Jensen’s foundation, Denmark and King Abdulaziz City for Science and Technology (09-BIO740-20), KSA. H. Saeed has received a PhD fellowship from the NovoNordisk Foundation, Denmark.

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Authors have no conflicting interests.

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Authors and Affiliations

  1. Endocrine Research Laboratory (KMEB), Department of Endocrinology and Metabolism, Odense University Hospital & University of Southern Denmark, Odense, Denmark

    Hamid Saeed, Hanna Taipaleenmäki, Abdullah M. Aldahmash, Basem M. Abdallah & Moustapha Kassem

  2. Department of Medical Biochemistry and Genetics, University of Turku, Turku, Finland

    Hanna Taipaleenmäki

  3. Faculty of Science, Helwan University, Cairo, Egypt

    Basem M. Abdallah

  4. Stem cell unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh, Saudi Arabia

    Abdullah M. Aldahmash & Moustapha Kassem

  5. Department of Endocrinology and Metabolism, Odense University Hospital, Kløvervægnet 6.4, DK-5000, Odense C, Denmark

    Moustapha Kassem

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  1. Hamid Saeed
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  2. Hanna Taipaleenmäki
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  3. Abdullah M. Aldahmash
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  4. Basem M. Abdallah
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  5. Moustapha Kassem
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Correspondence to Moustapha Kassem.

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Saeed, H., Taipaleenmäki, H., Aldahmash, A.M. et al. Mouse Embryonic Fibroblasts (MEF) Exhibit a Similar but not Identical Phenotype to Bone Marrow Stromal Stem Cells (BMSC). Stem Cell Rev and Rep 8, 318–328 (2012). https://doi.org/10.1007/s12015-011-9315-x

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