Inhibition of p53-p21 pathway promotes the differentiation of rat bone marrow mesenchymal stem cells into cardiomyocytes

Abstract

P53 is shown recently to play an important role in the proliferation and differentiation of bone marrow mesenchymal stem cells (BMMSCs). In this study, by inhibiting p53-p21 pathway with p53 inhibitor (p-fifty three inhibitor-alpha, PFT-α), we investigated the resulting effects on the differentiation of rat BMMSCs into cardiomyocyte-like cells. BMMSCs were isolated from bone marrow of SD rats by density gradient centrifugation. The fourth passage cells were divided into four groups: control group, PFT-α group, 5-AZA group, and PFT-α + 5-AZA group. The purified BMMSCs were identified by surface antigens and the proliferation and apoptosis of BMMSCs were examined by MTT and flow cytometry analysis. The expression of cTnI and CX-43 in BMMSCs after induction was detected by immunofluorescence and that of cTnI, p53, and p21 was detected by western blot. Our results demonstrated that PFT-α at 20 μmol/l significantly reduced the apoptosis and promoted the proliferation of BMMSCs, and induced BMMSCs to differentiate into cardiomyocyte-like cells. In conclusion, these data open up new possibility of modulating p53-p21 pathway for directed differentiation of BMMSCs into cardiomyocytes, which will be valuable for cardiovascular regenerative medicine.

Immunomodulatory effects of bee pollen on doxorubicin‐induced bone marrow/spleen immunosuppression in rat

“…The following primary antibodies were used: p‐53 Cat No: MAB1355; caspase‐3 Cat No: AF‐605‐NA, p‐21 Cat No: AF1047, and β‐actin Cat No: MAB8929 R&D Inc., USA). Immunoreactivity signals were detected by Odysseys Infrared Imager (Li‐COR Biosciences, Lincoln, NE, USA) device and analyzed with image J software (Yan et al., 2011).…”

Combination of BMP‐2 and 5‐AZA is advantageous in rat bone marrow‐derived mesenchymal stem cells differentiation into cardiomyocytes

“…BMMSCs are a promising option for stem cell transplantation therapy or tissue engineering (Yan et al, ; Xing et al, ). There are two ways of using BMMSCs for coronary heart disease therapy.…”

“…By 1 week later the cells had proliferated rapidly and formed "whirlpool-like" structures. After passage, cells were larger and more disordered, with a larger nucleocytoplasmic ratio ( Figure 1A; Yan et al, 2011;Xing et al, 2012). To quantify them, fourth passage BMMSCs were analysed by flow cytometry.…”

Mesenchymal Stem Cells and Cardiovascular Disease: A Bench to Bedside Roadmap

“…Cardiomyocyte differentiation has proved more problematic, as either demethylating agents have been employed [26], or it has been inefficient and incomplete [27, 28]. In contrast, the cardiac potential of ADSC is better documented in vitro , showing their capacity to give rise to CM, either by the use of DMSO [29] or CM extracts [30].…”