Transfection of STAT3 overexpression plasmid mediated through recombinant lentivirus promotes differentiation of bone marrow mesenchymal stem cells into neural cells in fetal rats with spina bifida aperta

Abstract

We investigated the influence of signal transducer and activator of transcription-3 (STAT3) on the spinal cord tissue grafts of rat fetuses with spina bifida aperta. In particular, we hoped to identify whether transfection of the STAT3 overexpression plasmid increases the survival of spinal cord transplantation in order to improve therapeutic efficacy. The fetal rat model of spina bifida aperta was established using retinoic acid and treated with a microsurgical injection of bone marrow mesenchymal stem cells (BMSCs). The animals were divided into either the blank control group, negative control group or the experimental group. The optical density (OD) value of BMSCs viability was determined using the Cell Counting Kit-8 (CCK-8). The expression of STAT3, phosphorylated STAT3 (pSTAT3), neural markers and apoptosis-related factors were evaluated using real-time PCR and Western blot. The OD value in the experimental group was highest at eight hours after transplantation using CCK-8. The expression of pSTAT3, glial fibrillary acidic protein, neuron-specific enolase, neurofilament and nestin in the experimental group was significantly higher compared to the blank control group and negative control group ( P <0.05). However, STAT3 expression in the experimental group was statistically significantly decreased ( P< 0.05). The relative expression of caspase-8 and bcl-2 in the experimental group were significantly lower compared to the blank control group and negative control group ( P <0.05). Transfection of the recombinant lentivirus-mediated STAT3 overexpression plasmid with BMSCs can help improve the efficiency of transforming into neural cells and provide new seed cells for the treatment of congenital spina bifida aperta.

MSC based gene delivery methods and strategies improve the therapeutic efficacy of neurological diseases

“…Hence, STAT3 overexpression in MSCs is a promising strategy for cell-based therapy. Consistent with this notion, transplanted BMSCs overexpressing STAT3 via recombinant lentivirus infection promoted the survival of fetal rats with spina bifida aperta (SBA), and this effect was associated with reduced BMSC apoptosis, enhanced BMSC survival, and increased neural marker expression [ 165 ]. Overexpression of the transcription factor nuclear receptor-related factor 1 (Nurr1), a member of the orphan nuclear receptor family implicated in dopaminergic neuron development and maturation, accelerated the dopaminergic differentiation of UC-MSCs [ 166 ] and BMSCs [ 167 ] and promoted the recovery of a rat model of Parkinson's disease (PD).…”

Transfection of Vascular Endothelial Growth Factor B into hypertrophic cardiomyocytes promotes angiogenesis by increasing endothelial cell VEGFR1 phosphorylation levels

“…Total RNA was extracted from cells and tissues using TRIzol reagent under RNase-free conditions, according to the manufacturer's protocol. All quali ed RNA was reverse transcribed using random primers and stem-loop primers in the RT Reagent Kit system [15]. β-Actin was used as the housekeeping gene.…”

Aging of mesenchymal stem cell: machinery, markers, and strategies of fighting

“…Though the JAK2/STAT3 pathway upregulated adipogenesis and restricted osteogenesis in a leptindependent manner, STAT3 may enhance MSCs' migration ability to increase their therapeutic efficacy [196]. It is suggested that the STAT3 pathway can be involved in inducing differentiation of BMSCs into neural cells [197] and have an antisenescence role [198]. Taken together, maintaining the correct function of STAT3 is an important issue for aging homeostasis in MSCs.…”