Adhesive Properties of the Hyaluronan Pericellular Coat in Hyaluronan Synthases Overexpressing Mesenchymal Stem Cells

被引:13
作者
Reiprich, Sebastian [1 ]
Hofbauer, Eva [1 ]
Kiderlen, Stefanie [2 ,3 ]
Clausen-Schaumann, Hauke [2 ,3 ]
Boecker, Wolfgang [1 ]
Aszodi, Attila [1 ]
Schoenitzer, Veronika [1 ]
机构
[1] Ludwig Maximilians Univ Munchen, Munich Univ Hosp, Expt Surg & Regenerat Med ExperiMed, Dept Gen Trauma & Reconstruct Surg, D-80336 Munich, Germany
[2] Munich Univ Appl Sci, Ctr Appl Tissue Engn & Regenerat Med, D-80533 Munich, Germany
[3] Ludwig Maximilians Univ Munchen, Ctr NanoSci, D-80799 Munich, Germany
关键词
hyaluronan; hyaluronan synthases; mesenchymal stem cells; early cell adhesion; stem cell niche; BONE-MARROW; MATRIX ADHESION; EXPRESSION; ISOFORMS; HAS1;
D O I
10.3390/ijms21113827
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Hyaluronan (HA), a natural component of the extracellular matrix, is supposed to have a regulatory function in the stem cell niche. Bone marrow-derived human mesenchymal stem cells (hMSCs) are known to express all three hyaluronan synthases (HASes), which are responsible for HA production. HA is extruded into the extracellular matrix, but also stays bound to the plasma membrane forming a pericellular coat, which plays a key role during early cell adhesion. Since HAS isoenzymes, HAS1, HAS2 and HAS3, produce HA with different molecular weights, a difference in their role for cell adhesion is expected. Here, we transduced the immortalized hMSC cell line SCP1 to constitutively express eGFP-tagged HASes (SCP1-HAS-eGFP) by lentiviral gene transfer. The overexpression of the HAS-eGFP was shown on RNA and protein levels, HA was determined by ELISA and the stained HA-coat was analyzed using confocal microscopy. Time-lapse microscopy, spreading assay and single cell force spectroscopy using atomic force microscopy were applied to characterize adhesion of the different HAS transduced SCP1 cells. We showed in this study that HAS3 overexpressing cells formed the thickest pericellular coat compared with control or HAS1 and HAS2 transduced cells. Furthermore, SCP1-HAS3-eGFP displayed faster and stronger adhesion compared to cells overexpressing the other synthases or control cells. We conclude that overexpression of HASes in hMSCs differentially modulates their initial adhesive interactions with the substrate. This observation might be helpful in regenerative medicine goals.
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页数:18
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