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Cell Surface Glycans as Markers of Shear Stimulated Stem Cells
Author(s) -
Weinstock Laura,
AdeniranCatlett Adedayo,
Bozal Fazli,
Murthy Shashi
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.890.14
Subject(s) - mesenchymal stem cell , chemistry , glycan , microbiology and biotechnology , stem cell , cell , biochemistry , biology , glycoprotein
Shear‐stress stimulation from perfusion through microfluidic tubing can induce adipogenesis in undifferentiated human mesenchymal stem cells (hMSCs). Increases in core‐fucosylated and sialylated biantennary N‐linked cell surface glycans have been shown to increase in adipogenically differentiated hMSCs as compared to their undifferentiated counterparts. Selecting for adipocyte‐associated glycans will allow for extracellular characterization of adipocytes, which can provide a more efficient identification method than measuring intracellular markers. We show that use of fluorescent microarray and immunocytochemical staining methods identify cell surface glycans associated with differentiation. We report here the use of lectins, rather than antibodies, to selectively bind glycans because the poor immunogenicity of glycans makes antibody production complicated and costly. The hMSCs were stained with fluorescent Lens culinaris Agglutinin (LCA), Wheat Germ Agglutinin (WGA), and Phaseolus vulgaris erythroagglutinin (PHA‐E) and imaged at days 0, 15, 25, and 32 after microfluidic passaging. Through fluorescent imaging we expect to see an increase of WGA and PHA‐E and a decrease of LCA in the shear‐stress stimulated, late stage cells. Rapid selection of adipocytes is expected to have important uses in reparative and cosmetic surgery involving autologous fat replacement.