Human induced pluripotent stem cell derived cardiac fibroblasts(hiPSC-CFs)play a critical role in modeling human cardiovascular diseases in vitro.However,current culture substrates used for hiPSC-CF differentiation an...Human induced pluripotent stem cell derived cardiac fibroblasts(hiPSC-CFs)play a critical role in modeling human cardiovascular diseases in vitro.However,current culture substrates used for hiPSC-CF differentiation and expansion,such as Matrigel and tissue culture plastic(TCPs),are tissue mismatched and may provide pathogenic cues.Here,we report that hiPSC-CFs differentiated on Matrigel and expanded on tissue culture plastic(M-TCP-iCFs)exhibit transcriptomic hallmarks of activated fibroblasts limiting their translational potential.To alleviate pathogenic activation of hiPSC-CFs,we utilized decellularized extracellular matrix derived from porcine heart extracellular matrix(HEM)to provide a biomimetic substrate for improving hiPSC-CF phenotypes.We show that hiPSC-CFs differentiated and expanded on HEM(HEM-iCFs)exhibited reduced expression of hallmark activated fibroblast markers versus M-TCP-iCFs while retaining their cardiac fibroblast phenotype.HEM-iCFs also maintained a reduction in expression of hallmark genes associated with pathogenic fibroblasts when seeded onto TCPs.Further,HEM-iCFs more homogenously integrated into an hiPSC-derived cardiac organoid model,resulting in improved cardiomyocyte sarcomere development.In conclusion,HEM provides an improved substrate for the differentiation and propagation of hiPSC-CFs for disease modeling.展开更多
基金funded by the National Institutes of Health(NIH)F31 HL154665,R01 HL133308,8P20 GM103444,R21 HL167211,R01 HL168255,U01 HL169361,R01 HL148059the NIH Cardiovascular Training Grant T32 HL007260+2 种基金the National Science Foundation(NSF)(CBET-1743346),the NSF Engineering Research Center for Cell Manufacturing Technologies(CMaT,NSF EEC-1648035)the VA Merit I01 BX005943-01A1supported by the grants,2021R1A2C3004262,2022M3A9B6082675,of the National Research Foundation of Korea(NRF)funded by the Korean government,the Ministry of Science and ICT(MSIT).
文摘Human induced pluripotent stem cell derived cardiac fibroblasts(hiPSC-CFs)play a critical role in modeling human cardiovascular diseases in vitro.However,current culture substrates used for hiPSC-CF differentiation and expansion,such as Matrigel and tissue culture plastic(TCPs),are tissue mismatched and may provide pathogenic cues.Here,we report that hiPSC-CFs differentiated on Matrigel and expanded on tissue culture plastic(M-TCP-iCFs)exhibit transcriptomic hallmarks of activated fibroblasts limiting their translational potential.To alleviate pathogenic activation of hiPSC-CFs,we utilized decellularized extracellular matrix derived from porcine heart extracellular matrix(HEM)to provide a biomimetic substrate for improving hiPSC-CF phenotypes.We show that hiPSC-CFs differentiated and expanded on HEM(HEM-iCFs)exhibited reduced expression of hallmark activated fibroblast markers versus M-TCP-iCFs while retaining their cardiac fibroblast phenotype.HEM-iCFs also maintained a reduction in expression of hallmark genes associated with pathogenic fibroblasts when seeded onto TCPs.Further,HEM-iCFs more homogenously integrated into an hiPSC-derived cardiac organoid model,resulting in improved cardiomyocyte sarcomere development.In conclusion,HEM provides an improved substrate for the differentiation and propagation of hiPSC-CFs for disease modeling.
