Baicalin Maintains Late-Stage Functional Cardiomyocytes in Embryoid Bodies Derived from Murine Embryonic Stem Cells

Abstract

Background/Aims: Low efficiency of cardiomyocyte (CM) differentiation from embryonic stem (ES) cells limits their therapeutic use. The objective of this study was to investigate the effect of baicalin, a natural flavonoid compound, on the in vitro cardiac differentiation of murine ES cells. Methods: The induction of ES cells into cardiac-like cells was performed by embryoid body (EB)-based differentiation method. The electrophysiological properties of the ES cell-derived CMs (ES-CMs) were measured by patch-clamp. The biomarkers of ES-CMs were determined by quantitative RT-PCR and immunofluorescence. Results: Continuous baicalin treatment decreased the size of EBs, and increased the proportion of α-actinin-positive CMs and transcript level of cardiac specific markers in beating EBs by inducing cell death of non-CMs. Baicalin increased the percentage of working ES-CMs which had typical responses to β-adrenergic and muscarinic stimulations. Conclusion: Baicalin maintains the late-stage functional CMs in EBs derived from murine ES cells. This study describes a new insight into the various biological effects of baicalin on cardiac differentiation of pluripotent stem cells.

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

“…The mES cell line D3 (ATCC, USA) and its transgenic cell line αPIG (clone 44) were cultured and differentiated into spontaneously beating CMs as previously described [3]. Briefly, the embryoid bodies (EBs) were generated using the hanging drop method in Iscove's Modified Dulbecco's Medium supplemented with 20 % fetal bovine serum, 1 % non-essential amino acids, 2 mM L-glutamine, 100 units/mL penicillin-streptomycin, and 0.1 mM β-mercaptoethanol.…”

“…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.…”

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

“…The differentiation of ES cells was induced by the generation of embryoid bodies (EBs) using the hanging drop method as previously described [11] with differentiation culture medium composed of Iscove's modified Dulbecco's medium (GIBCO), 20% FBS, 0.1 mmol/L β-mercaptoethanol, 1% NEAA, 2 mmol/L L-glutamine, 100 units/mL penicillin-streptomycin. Puerarin (1 µmol/L, 10 µmol/L, 100 µmol/L, 1000 µmol/L, Drug Biology Product Examination Bureau, Beijing, China) was added from day 0 to day 16.…”

“…So far, proteins from TGFβ, BMP, and Wnt families [4,5,6], cytokines [7], natural or synthetic chemical compounds [3,8,9,10,11]; and several transgenic ES cell systems [12] have been applied to selectively enhance the efficiency of cardiac differentiation from ES cells. Among these reports, retinoic acid and transgenic ES cell system are highlighted by their effects on ventricular specialization of ES cells.…”

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.…”