論文 - 詳細
| RRC ID | 59745 |
|---|---|
| 著者 | Aoyama J, Homma K, Tanabe N, Usui S, Miyagi Y, Matsuura K, Kaneda M, Nitta T. |
| タイトル | Spatiotemporal imaging documented the maturation of the cardiomyocytes from human induced pluripotent stem cells. |
| ジャーナル | J Thorac Cardiovasc Surg |
| Abstract |
OBJECTIVES:Cardiomyocytes derived from human induced pluripotent stem cells are a promising source of cells for regenerative medicine. However, contractions in such derived cardiomyocytes are often irregular and asynchronous, especially at early stages of differentiation. This study aimed to determine the differentiation stage of initiation of synchronized and regular contractions, using spatiotemporal imaging and physiological and genetic analyses. METHODS:Knock-in human induced pluripotent stem cell lines were established with clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeats-associated protein 9 to analyze cardiac and pacemaker cell maturation. Time-frequency analysis and Ca2+ imaging were performed, and the expression of related proteins and specific cardiac/pacemaker mRNAs in contracting embryoid bodies was analyzed at various differentiation stages. RESULTS:Time-frequency analysis and Ca2+ imaging revealed irregular, asynchronous contractions at the early stage of differentiation with altered electrophysiological properties upon differentiation. Genes associated with electrophysiological properties were upregulated after 70 days of culturing in differentiation media, whereas pacemaker genes were initially upregulated during the early stage and downregulated at the later stage. CONCLUSIONS:A differentiation period >70 days is required for adequate development of cardiac elements including ion channels and gap junctions and for sarcomere maturation. |
| 巻・号 | 159(6) |
| ページ | 2260-2271.e7 |
| 公開日 | 2020-6-1 |
| DOI | 10.1016/j.jtcvs.2019.06.060 |
| PII | S0022-5223(19)31358-3 |
| PMID | 31409490 |
| MeSH | Biological Clocks* / genetics Calcium Signaling* / genetics Cell Differentiation* / genetics Cell Line Gene Expression Regulation Gene Knock-In Techniques Humans Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels / genetics Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels / metabolism Induced Pluripotent Stem Cells / metabolism Induced Pluripotent Stem Cells / physiology* Muscle Proteins / genetics Muscle Proteins / metabolism Myocardial Contraction* / genetics Myocytes, Cardiac / metabolism Myocytes, Cardiac / physiology* Phenotype Potassium Channels / genetics Potassium Channels / metabolism T-Box Domain Proteins / genetics T-Box Domain Proteins / metabolism Time Factors |
| IF | 4.451 |
| 引用数 | 0 |
| リソース情報 | |
| ヒト・動物細胞 | 454E2(HPS0077) |