Article Text

GW24-e2201 Role of cardiotrophin-1 on cardiac differentiation of mouse induced pluripotent stem cells
  1. Liu Tong1,
  2. Cao Feng1
  1. 1Department of Cardiology, Xijing Hospital, Fourth Military Medical; Development Center for Tissue Engineering, Fourth Military Medical University, Xi’an, Shaanxi, China


Objectives Inducedpluripotent stem cell (iPSC), which demonstrates the stem cell characteristics like clonogenicity, self-renewal and multipotency, could differentiate into all three germ layer cell types including cardiomyocytes. The cytokinecardiotrophin-1, a member of IL-6 family, is highly expressed in the primitive hearttube during the embryonic cardiogenesis. This study aimed to investigate the role of cardiotrophin-1 (CT-1) on cardiac differentiation of mouse iPSC.

Methods MiPSC is cultured according to Yamanaka’s protocol. The differentiation of miPSC was initiated by embryoid bodies formation with classic hanging drop method. CT-1(1 μmol/ml) and vehicle control were administrated at day 3 of the differentiation process. Cells were harvested at day 4, 7, 10 and 14 after differentiation to examine the embryonic and cardiac specific cell markers expression by real-time PCR, immunofluorescence staining, flow cytometry analysis and ultra-microstructure changes by transmission electron microscope.

Results The iPS clone was in compactly oval or round shape with strong three-dimensional sense and distinct boundaries, and was Oct-4 positive in IF staining. The gene expressions of various cardiac specific cell markers in CT-1 group were significantly increased at different time points: Flk-1was increased 1.61 ± 0.03 folds at day 7 and 2.01 ± 0.01 folds at day 10, respectively (P < 0.05). Nkx2.5 was increased 2.08 ± 0.08 folds at day 7, 2.12 ± 0.15 folds at day 10 and 1.99 ± 0.06 folds at day 14, respectively (P < 0.01). Tbx5 was increased 1.61 ± 0.05 folds at day 7 and 1.86 ± 0.04 at day 10, respectively (P < 0.05). Furthermore, cTnT was increased1.75 ± 0.04 folds at day 10 and 1.78 ± 0.05 folds at day 14, respectively (P < 0.05). Flow cytometry analysis showed that cTnI positive cells were significantly increased in the CT-1 group compared to the vehicle group (56.4% vs 28.5%, P<0.05). More organised myofibril, intactmitochondria structure and well-formed cell junction were observed in the CT-1 group as compared with control.

Conclusions In the present study, we determined the role of cardiotrophin-1 on cardiac differentiation of miPSCs and assessed its effect at both molecular and cellular levels. Up-regulation of several cardiac mesodermal/progenitor markers such as Flk-1, Nkx2.5 and Tbx5 were observed in CT-1 group, indicating its enhancement of early cardiac differentiation transcriptional activities. These findings support the role of CT-1in the early stage of embryonic cardiogenesis. In addition, earlier appearance of beating EBs and more mature ultrastructure of iPS-derived cardiomyocytes (iPS-CMs) indicate CT-1 could enhance the maturity of iPS-CMs. In conclusion, CT-1 significantly promotes the cardiac differentiation of miPSCs as well asiPS-CM maturity and these findings provide a potent evidence for iPSC-based cardiac cell therapy of myocardial infarction or congestive heart failure.

Statistics from

Request permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.