Abstract
Adult Mesenchymal Stem Cells (MSC) are cells that can be defined as multipotent cells able to differentiate into diverse lineages, under appropriate conditions. These cells have been widely used in regenerative medicine, both in preclinical and clinical settings. Initially discovered in bone marrow, MSC can now be isolated from a wide spectrum of adult and foetal tissues. Studies to evaluate the therapeutic potential of these cells are based on their ability to arrive to damaged tissues. In this paper we have done a comparative study analyzing proliferation, surface markers and OCT4, SOX9, RUNX2, PPARG genes expression in MSC cells from Bone marrow (BMMSC) and Adipose tissue (ASC). We also analyzed the role of Stem Cell Factor (SCF) on MSC proliferation and on ASCs metalloproteinases MMP-2, MMP-9 secretion. Healthy dogs were used as BMMSC donors, and ASC were collected from omentum during elective ovariohysterectomy surgery. Both cell types were cultured in IMDM medium with or without SCF, 10% Dog Serum (DS), and incubated at 38 °C with 5% CO2. Growth of BMMSCs and ASCs was exponential until 25–30 days. Flow citometry of MSCs revealed positive results for CD90 and negative for CD34, CD45 and MCH-II. Genes were evaluated by RT-PCR and metalloproteinases by zymografy. Our findings indicate morphological and immunological similarities as well as expression of genes from both origins on analyzed cells. Furthermore, SCF did not affect proliferation of MSCs, however it up-regulated MMP-2 and MMP-9 secretion in ASCs. These results suggest that metalloproteinases are possibly essential molecules pivoting migration.
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Acknowledgements
We are very thankful to Dra. Carmen Fajardo for her scientific assistance. We thank Flow Cytometry, Genomics and Proteomics UCM facilities, for their technical support. Leticia G. León care Marie Curie grant. Authors would like to thank Elisabeth Salva for their technical assistance. Some studies reported in this manuscript were supported by a grant from the Deutsche José Carreras Leukämie Stiftung e.V. (R01/R13.)
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Standard phenotype from bone marrow BMMSCs and adipose tissue ASCs was confirmed.
We verified OCT4, SOX9, RUNX2, and PPARG genes expression in MSC from both origins.
ASCs constitutively express MMP-2 and MMP-9, being up-regulated by SCF.
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Enciso, N., Ostronoff, L.L.K., Mejías, G. et al. Stem cell factor supports migration in canine mesenchymal stem cells. Vet Res Commun 42, 29–38 (2018). https://doi.org/10.1007/s11259-017-9705-x
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DOI: https://doi.org/10.1007/s11259-017-9705-x