Generation and Characterization of Patient-Specific iPSC Model for Cardiovascular Disease

Yee Ki Lee; X Ran; K W H Lai; V Y M Lau; D C W Siu; H F Tse

doi:10.1007/7651_2015_273

Generation and Characterization of Patient-Specific iPSC Model for Cardiovascular Disease

Methods Mol Biol. 2016:1353:191-213. doi: 10.1007/7651_2015_273.

Authors

Yee Ki Lee¹, X Ran¹, K W H Lai¹, V Y M Lau¹, D C W Siu^{1

2

3

4}, H F Tse^{5

6

7

8

9}

Affiliations

¹ Cardiology Division, Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
² Research Center of Heart, Brain, Hormone and Healthy Aging, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China.
³ Shenzhen Institutes of Research and Innovation, University of Hong Kong, Hong Kong, China.
⁴ Division of Cardiology, Queen Mary Hospital, Hong Kong, China.
⁵ Cardiology Division, Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. hftse@hku.hk.
⁶ Research Center of Heart, Brain, Hormone and Healthy Aging, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China. hftse@hku.hk.
⁷ Shenzhen Institutes of Research and Innovation, University of Hong Kong, Hong Kong, China. hftse@hku.hk.
⁸ Hong Kong-Guangdong Joint Laboratory on Stem Cell and Regenerative Medicine, Hong Kong, China. hftse@hku.hk.
⁹ Division of Cardiology, Queen Mary Hospital, Hong Kong, China. hftse@hku.hk.

PMID: 26126449
DOI: 10.1007/7651_2015_273

Abstract

Advances in differentiation of cardiomyocytes from human induced pluripotent stem cell (hiPSC) were emerged as a tool for modeling of cardiovascular disease that recapitulates the phenotype for the purpose of drug screening, biomarker discovery, and testing of single-nucleotide polymorphism (SNP) as a modifier for disease stratification. Here, we describe the (1) retroviral reprogramming strategies in the generation of human iPSC, (2) methodology in characterization of iPSC in order to identify the stem cell clones with the best quality, and (3) protocol of cardiac differentiation by modulation of Wnt signaling and β-catenin pathway.

Keywords: Cardiac differentiation; Cardiovascular disease modeling; Human induced pluripotent stem cell (hiPSC); Retroviral reprogramming; Stem cell characterization.

MeSH terms

Amides / pharmacology
Animals
Biomarkers / metabolism
Cell Culture Techniques / methods*
Cell Differentiation
Cellular Reprogramming*
Collagen / chemistry
Drug Combinations
Embryo, Mammalian
Embryoid Bodies / cytology*
Embryoid Bodies / drug effects
Embryoid Bodies / metabolism
Enzyme Inhibitors / pharmacology
Feeder Cells / cytology
Fibroblasts / cytology
Gene Expression
Genetic Vectors
Humans
Induced Pluripotent Stem Cells / cytology*
Induced Pluripotent Stem Cells / drug effects
Induced Pluripotent Stem Cells / metabolism
Intercellular Signaling Peptides and Proteins / pharmacology
Laminin / chemistry
Mice
Myocytes, Cardiac / cytology*
Myocytes, Cardiac / drug effects
Myocytes, Cardiac / metabolism
Primary Cell Culture
Proteoglycans / chemistry
Pyridines / pharmacology
Pyrimidines / pharmacology
Retroviridae / genetics
Teratoma / genetics
Teratoma / metabolism
Teratoma / pathology
Wnt Signaling Pathway
beta Catenin / genetics
beta Catenin / metabolism

Substances

Amides
Biomarkers
CTNNB1 protein, human
Chir 99021
Drug Combinations
Enzyme Inhibitors
Intercellular Signaling Peptides and Proteins
Laminin
Proteoglycans
Pyridines
Pyrimidines
beta Catenin
matrigel
Y 27632
Collagen