In vitro analysis of primary isolated adult cardiomyocyte physiological processes often involves optical imaging of dye-loaded cells on a glass substrate. However, when exposed to rapid solution changes, primary cardiomyocytes often move to compromise quantitative measures. Improved immobilization of cells to glass would permit higher throughput assays. Here, we engineer the peripheral membrane of cardiomyocytes with biotin to anchor cardiomyocytes to borosilicate glass coverslips functionalized with streptavidin. We use a rat cardiac myoblast cell line to determine general relationships between processing conditions, ligand density on the cell and the glass substrate, cellular function, and cell retention under shear flow. Use of the streptavidin-biotin system allows for more than 80% retention of cardiac myoblasts under conventional rinsing procedures, while unmodified cells are largely rinsed away. The adhesion system enables the in-field retention of cardiac cells during rapid fluid changes using traditional pipetting or a modern microfluidic system at a flow rate of 160 mL/min. Under fluid flow, the surface-engineered primary adult cardiomyocytes are retained in the field of view of the microscope, while unmodified cells are rinsed away. Importantly, the engineered cardiomyocytes are functional following adhesion to the glass substrate, where contractions are readily observed. When applying this adhesion system to cardiomyocyte functional analysis, we measure calcium release transients by caffeine induction at an 80% success rate compared to 20% without surface engineering.
|Number of pages||8|
|State||Published - Jul 13 2021|
Bibliographical noteFunding Information:
This work was partially supported by the NIH under R01 HL127682, the National Science Foundation under Award CBET-1351531, and the American Heart Association grant # 18IPA34170059/Berron&Abdel-Latif/2018. Dr. A.A.-L. is supported by the NIH grant R01 HL138488. Dr. J.S. is supported by the NIH grant R01 HL131782. The project described was also supported by the NIH National Center for Advancing Translational Sciences through grant number TL1TR001997.
© 2021 The Authors. American Chemical Society.
ASJC Scopus subject areas
- Chemistry (all)
- Chemical Engineering (all)