Effects of Puerarin on Cardiac Differentiation and Ventricular Specialization of Murine Embryonic Stem Cells

Abstract

Aims: It is important to screen and identify chemical compounds to improve the efficiency of cardiac differentiation and specialization of embryonic stem (ES) cells. The objective of this study was to investigate the effect of puerarin, a natural phytoestrogen, on the in vitro cardiac differentiation and ventricular specialization of murine ES cells. Methods: Cardiac differentiation of murine ES cells was performed by embryoid body (EB)-based differentiation method. Quantitative RT-PCR, flow cytometry and immunofluorescence were employed to identify cardiomyocytes (CMs) derived from murine ES cells (mES-CMs). Patch clamp was used to study the electrophysiological properties of CMs. Results: We found that continuous puerarin treatment significantly increased the population of ES-CMs which express typical cardiac markers and are electrophysiological intact. Puerarin treatment shifted the cardiac phenotype from pacemaker-like cells to ventricular-like cells, which were Mlc2v-positive and present typical ventricular-like AP. Puerarin up-regulated transcripts involved in cardiac differentiation and ventricular specialization of ES cells. Conclusion: Our results suggest that puerarin promotes cardiac differentiation, and significantly enhances the specialization of mES cells into ventricular-like CMs. Puerarin may be used to increase the yield of ventricular mES-CMs during in vitro differentiation.

Puerarin Facilitates T-Tubule Development of Murine Embryonic Stem Cell-Derived Cardiomyocytes

“…Puerarin (100 μM, National Institutes For Food and Drug Control, China), which dissolved in 0.05 % dimethyl sulfoxide (DMSO, Sigma, USA), was applied from day 0 to day 20 as previously described [4]. EBs cultured with differentiation medium containing 0.05 % DMSO were severed as controls.…”

“…Our previous study found that puerarin enhances the cardiac differentiation and ventricular specialization of mES cells [4]. In this study, we tested the hypothesis that structural organization of mES-CMs would be improved by long-term application of puerarin throughout cardiac differentiation in vitro .…”

“…This strategy is transplanting suitable cells into the infarcted area of heart which can functionally integrate into the host tissue, and thus rescuing cardiac function [1]. Embryonic stem (ES) cells generated from blastocysts inner cell mass can differentiate into beating cardiomyocytes (CMs) in vitro , which have been widely accepted as a promising source for cell replacement therapy [1,2,3,4]. The typical structure of adult CMs is characterized by bundles of parallel myofilaments, sarcomeres with Z-disc, A-, I-, and H-band as well as the M-line, which are the structural basis of cardiac muscle contraction.…”

Puerarin Enhances Ca2+ Reuptake and Ca2+ Content of Sarcoplasmic Reticulum in Murine Embryonic Stem Cell-Derived Cardiomyocytes via Upregulation of SERCA2a

“…Accumulated studies have shown that the PI3K/Akt, JAK/STAT, JNK, ERK1/2 and MAPK signaling pathways mediate the bio-effects of puerarin on various types of cells, i.e CMs [2, 17, 20], astrocytes [23], neurons [24], and retinal ganglion cells [10]. Our previous research found that puerarin could enhance the specialization of mES cells into ventricular-like CMs [25], inhibits the proliferation of mES-CMs [2], while improves the t-tubule development of mES-CMs [4], suggesting an ameliorated excitation-contraction coupling of these cells in the presence of puerarin. However, it is still uncertain whether puerarin has effect on the Ca 2+ handling maturation of ES-CMs.…”

Traditional Chinese Medicine Baicalin Suppresses mESCs Proliferation through Inhibition of miR-294 Expression

“…These active components regulate either the transcription factors [13] or the cell cycle-related genes thereby affecting stem cell differentiation, proliferation and apoptosis. Our previous data have revealed that TCMs baicalin, puerarin and ginsenoside Rg1 influence the cardiac differentiation of mESCs [14,15,16] by changing the expression of the cardiac-specific transcriptional factors. Whether they could also exert effects on mESCs proliferation by modifying the expression of mESC-miRNAs remains to be determined.…”