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    • The following are the supplementary data related

    2018-10-20

    The following are the supplementary data related to this article.
    Author contribution
    Acknowledgment We thank Dr. Donald Vander Griend for providing CD133 cDNA, Dr. Oh-Joon Kwon and Ms. Li Zhang for technical help, the technical support by the Cytometry and Cell Sorting Core at Baylor College of Medicine with funding from the NIH (P30 AI036211, P30 CA125123, and S10 RR024574) and the expert assistance of Joel M. Sederstrom. This work is supported by NIHR01 DK092202 (L.X.) and NIHP30 CA125123 (the Cancer Center Shared Resources Grant).
    Introduction Adherent, fibroblastic molarity conversion from different tissues (e.g., from bone marrow, adipose tissue, umbilical cord blood, placenta, Wharton jelly, etc.) are thought to contain subsets of tissue-specific stem/progenitor cells (often called mesenchymal stem cells). These tissue-specific stem/progenitor cells share many biological features. However, they also display differences in molecular phenotype, growth rate, and their ability to differentiate into various phenotypes (Kern et al., 2006; Al-Nbaheen et al., 2013; Choudhery et al., 2013). Calcium is a ubiquitous intracellular messenger that is a key regulator of the cell cycle, particularly during stem cell proliferation and modification. The Ca signaling pathways have been studied in a variety of stem cell types including embryonic (Forostyak et al., 2013; Viero et al., 2014), fetal (Cocks et al., 2013) and adult stromal cells (Resende et al., 2010; Zippel et al., 2012; Kotova et al., 2014; Forostyak et al., 2016). Bone marrow stromal cells (BMSCs) have been shown to express L-type Ca channels (Heubach et al., 2004; Li et al., 2006; Wen et al., 2012), glutamate receptors (Fox et al., 2010) and have been reported to generate spontaneous inositol 1,4,5-triphosphate (InsP3)-dependent Ca oscillations (Kawano et al., 2002, 2003). Adipose tissue-derived stromal cells (ADSCs), were found to express adrenoceptors, InsP3 receptors (InsP3Rs), purinoceptors and were reported to generate Ca-induced Ca release (Kotova et al., 2014). BMSCs have also been shown to express specific K+ channels including Ca-activated K+ channels (IKCa), delayed rectifier K+ current (IKDR), and transient outward K+ current (Ito) (Li et al., 2006). In human BMSCs in basal conditions (bBMSCs) large conductance voltage- and Ca-activated K+ channels have been identified (Heubach et al., 2004). To the best of our knowledge, an in depth analysis of ion channels and receptors in ADSCs and BMSCs that have been harvested under the same environmental conditions has not been performed. In this study, we compared the functional properties of these two types of cells in basal conditions (bADSCs and bBMSCs) and after their modification (mADSCs and mADSC) induced by switching to a medium containing factors known to alter their characteristics.
    Experimental procedures
    Results
    Discussion In this study, we investigated Ca signaling and electrophysiological properties of rat stromal cells obtained from adipose tissue and bone marrow. We analyzed the changes of functional properties under the same environmental conditions upon medium modification that were previously reported to elicit changes in ADSCs (Willingham and Pastan, 1975). We demonstrated that after medium modification, both ADSCs and BMSCs undergo a significant change in their membrane properties and in the expression of channels and receptors associated with generation of Ca signals (Fig. 7). The resting [Ca]i level in mBMSCs significantly decreased after pre-differentiation and became almost the same as in ADSCs. The resting [Ca]i level in ADSCs remained unchanged after medium modification.
    Conclusions Together, as briefly summarized in Fig. 7, our results suggest that the Ca signaling, ion channel and ionotropic receptor expression profile of adult tissue-specific stem/progenitor cells depends not only on the culture conditions but also on the source from which the cells were isolated. The fate and functional properties of differentiated cells are driven by intrinsic mechanisms, which are independent of the culture protocol. Identifying their Ca signaling, ion channel and ionotropic receptor expression profile as a function of external changes in their environment is essential to better utilize these cells in tissue engineering and regenerative medicine.