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Efficient differentiation of human pluripotent stem cells into cardiomyocytes on cell sorting thermoresponsive surface.

Biomaterials (2020-05-26)
Tzu-Cheng Sung, Huan Chiao Su, Qing-Dong Ling, S Suresh Kumar, Yung Chang, Shih-Tien Hsu, Akon Higuchi
ABSTRACT

The current differentiation process of human pluripotent stem cells (hPSCs) into cardiomyocytes to enhance the purity of hPSC-derived cardiomyocytes requires some purification processes, which are laborious processes. We developed cell sorting plates, which are prepared from coating thermoresponsive poly(N-isopropylacrylamide) and extracellular matrix proteins. After hPSCs were induced into cardiomyocytes on the thermoresponsive surface coated with laminin-521 for 15 days, the temperature of the cell culture plates was decreased to 8-9 °C to detach the cells partially from the thermoresponsive surface. The detached cells exhibited a higher cardiomyocyte marker of cTnT than the remaining cells on the thermoresponsive surface as well as the cardiomyocytes after purification using conventional cell selection. The detached cells expressed several cardiomyocyte markers, such as α-actinin, MLC2a and NKX2.5. This study suggested that the purification of hPSC-derived cardiomyocytes using cell sorting plates with the thermoresponsive surface is a promising method for the purification of hPSC-derived cardiomyocytes without conventional laborious processes.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
CHIR99021, ≥98% (HPLC)
Sigma-Aldrich
Monoclonal Anti-α-Actinin (Sarcomeric) antibody produced in mouse, clone EA-53, ascites fluid
Sigma-Aldrich
IWR-1, ≥98% (HPLC)
Supelco
3,5,6-Trichloro-2-pyridinol, PESTANAL®, analytical standard