Objective To investigate whether cardiac tissue extracts from rats could mimic the cardiac microenvironment and act as a natural inducer in promoting the differentiation of bone marrow stromal cells (BMSCs) into car...Objective To investigate whether cardiac tissue extracts from rats could mimic the cardiac microenvironment and act as a natural inducer in promoting the differentiation of bone marrow stromal cells (BMSCs) into cardiomyocytes. Methods Three kinds of tissue extract or cell lysate [infarcted myocardial tissue extract (IMTE), normal myocardial tissue extract (NMTE) and cultured neonatal myocardial lysate (NML)] were employed to induce BMSCs into cardiomyocyte-like cells. The cells were harvested at each time point for reverse transcription-polymerase chain reaction (RT-PCR) detection, immunocytochemical analysis, and transmission electron microscopy. Results After a 7-day induction, BMSCs were enlarged and polygonal in morphology. Myofilaments, striated sarcomeres, Z-lines, and more mitochondia were observed under transmission electron microscope. Elevated expression levels of cardiac-specific genes and proteins were also confirmed by RT-PCR and immunocytochemistry. Moreover, IMTE showed a greater capacity of differentiating BMSCs into cardiomyocyte-like cells. Conclusions Cardiac tissue extracts, especially IMTE, can effectively differentiate BMSCs into cardiomyocyte-like cells.展开更多
Background:To understand the relationship between myocardial contractility and ex-ternal stimuli,detecting ex vivo myocardial contractility is necessary.Methods:We elaborated a method for contractility detection of is...Background:To understand the relationship between myocardial contractility and ex-ternal stimuli,detecting ex vivo myocardial contractility is necessary.Methods:We elaborated a method for contractility detection of isolated C57 mouse papillary muscle using Myostation-Intact system under different frequencies,volt-ages,and calcium concentrations.Results:The results indicated that the basal contractility of the papillary muscle was 0.27±0.03 mN at 10 V,500-ms pulse duration,and 1 Hz.From 0.1 to 1.0 Hz,con-tractility decreased with an increase in frequency(0.45±0.11-0.10±0.02 mN).The voltage-initiated muscle contractility varied from 3 to 6 V,and the contractility gradu-ally increased as the voltage increased from 6 to 10 V(0.14±0.02-0.28±0.03 mN).Moreover,the muscle contractility increased when the calcium concentration was increased from 1.5 to 3 mM(0.45±0.17-1.11±0.05 mN);however,the contractility stopped increasing even when the concentration was increased to 7.5 mM(1.02±0.23 mN).Conclusions:Our method guaranteed the survivability of papillary muscle ex vivo and provided instructions for Myostation-Intact users for isolated muscle contractility investigations.展开更多
The RNA editing tool CRISPR-CasRx has provided a platform for a range of transcriptome analysis tools and therapeutic approaches with its broad efficacy and high specificity.To enable the application of CasRx in vivo,...The RNA editing tool CRISPR-CasRx has provided a platform for a range of transcriptome analysis tools and therapeutic approaches with its broad efficacy and high specificity.To enable the application of CasRx in vivo,we established a Credependent CasRx knock-in mouse.Using these mice,we specifically knocked down the expression of Meis1 and Hoxb13 in cardiomyocytes,which induced cardiac regeneration after myocardial infarction.We also knocked down the lnc RNA Mhrt in cardiomyocytes with the CasRx knock-in mice,causing hypertrophic cardiomyopathy.In summary,we generated a Credependent CasRx knock-in mouse that can efficiently knock down coding gene and lnc RNA expression in specific somatic cells.This in vivo CRISPR-CasRx system is promising for gene function research and disease modeling.展开更多
基金This work was supported by the National Natural Science Foundation of China (No. 30570722)
文摘Objective To investigate whether cardiac tissue extracts from rats could mimic the cardiac microenvironment and act as a natural inducer in promoting the differentiation of bone marrow stromal cells (BMSCs) into cardiomyocytes. Methods Three kinds of tissue extract or cell lysate [infarcted myocardial tissue extract (IMTE), normal myocardial tissue extract (NMTE) and cultured neonatal myocardial lysate (NML)] were employed to induce BMSCs into cardiomyocyte-like cells. The cells were harvested at each time point for reverse transcription-polymerase chain reaction (RT-PCR) detection, immunocytochemical analysis, and transmission electron microscopy. Results After a 7-day induction, BMSCs were enlarged and polygonal in morphology. Myofilaments, striated sarcomeres, Z-lines, and more mitochondia were observed under transmission electron microscope. Elevated expression levels of cardiac-specific genes and proteins were also confirmed by RT-PCR and immunocytochemistry. Moreover, IMTE showed a greater capacity of differentiating BMSCs into cardiomyocyte-like cells. Conclusions Cardiac tissue extracts, especially IMTE, can effectively differentiate BMSCs into cardiomyocyte-like cells.
基金Specialized Project of Fuwai Hospital,Grant/Award Number:2022-FWTS07Shenzhen Sanming Project of Medicine,Grant/Award Number:2016-SZZF02+1 种基金National Natural Science Foundation of China,Grant/Award Number:81900343CAMS Innovation Fund for Medical Sciences,Grant/Award Number:CIFMS,2021-I2M-C&T-A-011。
文摘Background:To understand the relationship between myocardial contractility and ex-ternal stimuli,detecting ex vivo myocardial contractility is necessary.Methods:We elaborated a method for contractility detection of isolated C57 mouse papillary muscle using Myostation-Intact system under different frequencies,volt-ages,and calcium concentrations.Results:The results indicated that the basal contractility of the papillary muscle was 0.27±0.03 mN at 10 V,500-ms pulse duration,and 1 Hz.From 0.1 to 1.0 Hz,con-tractility decreased with an increase in frequency(0.45±0.11-0.10±0.02 mN).The voltage-initiated muscle contractility varied from 3 to 6 V,and the contractility gradu-ally increased as the voltage increased from 6 to 10 V(0.14±0.02-0.28±0.03 mN).Moreover,the muscle contractility increased when the calcium concentration was increased from 1.5 to 3 mM(0.45±0.17-1.11±0.05 mN);however,the contractility stopped increasing even when the concentration was increased to 7.5 mM(1.02±0.23 mN).Conclusions:Our method guaranteed the survivability of papillary muscle ex vivo and provided instructions for Myostation-Intact users for isolated muscle contractility investigations.
基金the National Key Research and Development Project of China(2019YFA0801500)Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences(CIFMS,2021-I2M-1-008)the National Natural Science Foundation of China(81770308,81900343)。
文摘The RNA editing tool CRISPR-CasRx has provided a platform for a range of transcriptome analysis tools and therapeutic approaches with its broad efficacy and high specificity.To enable the application of CasRx in vivo,we established a Credependent CasRx knock-in mouse.Using these mice,we specifically knocked down the expression of Meis1 and Hoxb13 in cardiomyocytes,which induced cardiac regeneration after myocardial infarction.We also knocked down the lnc RNA Mhrt in cardiomyocytes with the CasRx knock-in mice,causing hypertrophic cardiomyopathy.In summary,we generated a Credependent CasRx knock-in mouse that can efficiently knock down coding gene and lnc RNA expression in specific somatic cells.This in vivo CRISPR-CasRx system is promising for gene function research and disease modeling.