TY - JOUR
T1 - Small molecule-mediated rapid maturation of human induced pluripotent stem cell-derived cardiomyocytes
AU - Chirico, Nino
AU - Kessler, Elise L
AU - Maas, Renée G C
AU - Fang, Juntao
AU - Qin, Jiabin
AU - Dokter, Inge
AU - Daniels, Mark
AU - Šarić, Tomo
AU - Neef, Klaus
AU - Buikema, Jan-Willem
AU - Lei, Zhiyong
AU - Doevendans, Pieter A
AU - Sluijter, Joost P G
AU - van Mil, Alain
N1 - Funding Information:
NC is supported by the Gravitation Program “Materials Driven Regeneration” by the Netherlands Organization for Scientific Research (RegmedXB #024.003.013) and the Marie Skłodowska-Curie Actions (Grant agreement RESCUE #801540). RM is supported by a grant of the PLN Foundation and HARVEY (18747 NWO OTP). AvM is supported by the EU-funded project BRAV3 (H2020, ID:874827). This work was supported by European Research Council (ERC) under the EVICARE Grant (number 725229) to JS.
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/12/27
Y1 - 2022/12/27
N2 - BACKGROUND: Human induced pluripotent stem cell (iPSC)-derived cardiomyocytes (iPSC-CMs) do not display all hallmarks of mature primary cardiomyocytes, especially the ability to use fatty acids (FA) as an energy source, containing high mitochondrial mass, presenting binucleation and increased DNA content per nuclei (polyploidism), and synchronized electrical conduction. This immaturity represents a bottleneck to their application in (1) disease modelling-as most cardiac (genetic) diseases have a middle-age onset-and (2) clinically relevant models, where integration and functional coupling are key. So far, several methods have been reported to enhance iPSC-CM maturation; however, these protocols are laborious, costly, and not easily scalable. Therefore, we developed a simple, low-cost, and rapid protocol to promote cardiomyocyte maturation using two small molecule activators of the peroxisome proliferator-activated receptor β/δ and gamma coactivator 1-alpha (PPAR/PGC-1α) pathway: asiatic acid (AA) and GW501516 (GW). METHODS AND RESULTS: Monolayers of iPSC-CMs were incubated with AA or GW every other day for ten days resulting in increased expression of FA metabolism-related genes and markers for mitochondrial activity. AA-treated iPSC-CMs responsiveness to the mitochondrial respiratory chain inhibitors increased and exhibited higher flexibility in substrate utilization. Additionally, structural maturity improved after treatment as demonstrated by an increase in mRNA expression of sarcomeric-related genes and higher nuclear polyploidy in AA-treated samples. Furthermore, treatment led to increased ion channel gene expression and protein levels.CONCLUSIONS: Collectively, we developed a fast, easy, and economical method to induce iPSC-CMs maturation via PPAR/PGC-1α activation. Treatment with AA or GW led to increased metabolic, structural, functional, and electrophysiological maturation, evaluated using a multiparametric quality assessment.
AB - BACKGROUND: Human induced pluripotent stem cell (iPSC)-derived cardiomyocytes (iPSC-CMs) do not display all hallmarks of mature primary cardiomyocytes, especially the ability to use fatty acids (FA) as an energy source, containing high mitochondrial mass, presenting binucleation and increased DNA content per nuclei (polyploidism), and synchronized electrical conduction. This immaturity represents a bottleneck to their application in (1) disease modelling-as most cardiac (genetic) diseases have a middle-age onset-and (2) clinically relevant models, where integration and functional coupling are key. So far, several methods have been reported to enhance iPSC-CM maturation; however, these protocols are laborious, costly, and not easily scalable. Therefore, we developed a simple, low-cost, and rapid protocol to promote cardiomyocyte maturation using two small molecule activators of the peroxisome proliferator-activated receptor β/δ and gamma coactivator 1-alpha (PPAR/PGC-1α) pathway: asiatic acid (AA) and GW501516 (GW). METHODS AND RESULTS: Monolayers of iPSC-CMs were incubated with AA or GW every other day for ten days resulting in increased expression of FA metabolism-related genes and markers for mitochondrial activity. AA-treated iPSC-CMs responsiveness to the mitochondrial respiratory chain inhibitors increased and exhibited higher flexibility in substrate utilization. Additionally, structural maturity improved after treatment as demonstrated by an increase in mRNA expression of sarcomeric-related genes and higher nuclear polyploidy in AA-treated samples. Furthermore, treatment led to increased ion channel gene expression and protein levels.CONCLUSIONS: Collectively, we developed a fast, easy, and economical method to induce iPSC-CMs maturation via PPAR/PGC-1α activation. Treatment with AA or GW led to increased metabolic, structural, functional, and electrophysiological maturation, evaluated using a multiparametric quality assessment.
KW - Asiatic acid
KW - GW501516
KW - Human induced pluripotent stem cell-derived cardiomyocyte
KW - Maturation
KW - PGC-1α
KW - PPAR
UR - http://www.scopus.com/inward/record.url?scp=85144749246&partnerID=8YFLogxK
U2 - 10.1186/s13287-022-03209-z
DO - 10.1186/s13287-022-03209-z
M3 - Article
C2 - 36575473
SN - 1757-6512
VL - 13
SP - 1
EP - 18
JO - Stem Cell Research & Therapy [E]
JF - Stem Cell Research & Therapy [E]
IS - 1
M1 - 531
ER -