Abstract 15234: A Novel Purification Method of Cardiomyocytes Derived From iPS Cells, Based on Their Affinity to Laminin-221 in Terms of Clinical Applications
Introduction: Although cardiomyocytes (CMs) derived from human-induced pluripotent stem cells (hiPSCs) have the potential to treat heart diseases, quality control of differentiated CMs, especially in terms of purification, is crucial for clinical application. We hypothesized that the ability of CMs to adhere to laminin (LM) - that is, the major extracellular matrix components in myocardium - may lead to new purification methods of hiPSCs-derived CMs.
Methods: (1) The expression pattern of integrins on the cell surface of differentiated CMs and undifferentiated hiPSCs were analyzed by quantitative PCR to check which type of LM was suitable for effective purification in CMs. (2) We compared various extracellular matrix-coated dishes to determine whether they had high affinity to CMs and could be purified by 30 min plating of CMs. (3) We implanted a post-purified CMs sheet (post group: n=5) and a pre-purified CMs sheet (pre group: n=5) to nude rat chronic infarction models and evaluated functional recovery compared with a sham-operated group (S group: n=6).
Results: (1) We found that hiPSCs-derived CMs expressed an approximately 10- to 40-fold increase in integrin subunits α1, α3, and α7, which have high affinity to LM-221, compared with undifferentiated hiPSCs (p<0.01), suggesting that LM-221 may be suitable for purifying differentiated CMs. (2) Flow cytometry analysis demonstrated that CMs were significantly purified by plating on LM-221-coated dishes, compared with pre-value (pre vs post purity: 50%±7% vs 63%±6%; p<0.05, n=7). In contrast, there was no significant difference in pre- and post-purity of CMs when plating on LM-411, -511, fibronectin-coated and uncoated dishes. (3) Four weeks after the transplantation, the ejection fraction was significantly higher in the post group compared with the S group and pre group when measured by echocardiography (post 56±5% vs pre 52±4% vs S 39±3%, p<0.01). Four weeks post-op, the expressions of mouse VEGF, SDF-1, and HGF mRNA levels in the ischemic area were increased in the post group compared with S groups (p<0.05).
Conclusion: We developed a novel purification system enabling the production of purified CMs, based on the affinity of CMs to LM-221, promising key clinical applications of such CMs for heart failure.
Author Disclosures: F. Ohashi: Employment; Significant; Terumo Corporation. S. Miyagawa: None. S. Fukushima: None. Y. Imanishi: None. S. Yoshida: None. A. Saito: None. S. Masuda: None. H. Iseoka: Employment; Significant; Terumo Corporation. E. Ito: None. T. Ishikawa: None. T. Sameshima: Employment; Significant; Terumo Corporation. K. Sekiguchi: None. Y. Sawa: None.
- © 2016 by American Heart Association, Inc.