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.

Mingyu Jiang,Mingyong Ren,Chunming Jiang,Yanbo Pan,Jiale Feng,Xu Liu,Yunpeng Hao,Rong Fu,Jicheng Dai

doi:10.18632/aging.203524

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.

Highlights

Neural tube defects (NTDs) in children can result in severe birth abnormalities
Cell Counting Kit-8 (CCK-8) assay for the assessment of effective transfection in the proliferative activity of bone marrow mesenchymal stem cells (BMSCs), and the signal transducer and activator of transcription-3 (STAT3) plasmid prior to transplantation
The STAT3 pathway controls a number of embryonic development pathways and is involved in aberrant embryonic development, which can be affected by valproic acid [16]

Summary

Introduction

Neural tube defects (NTDs) in children can result in severe birth abnormalities. The primary mechanism of NTDs is currently considered to be aberrant neural tube closure. Congenital spina bifida aperta (SBA) is the most prevalent of many kinds of NTDs [1]. Apoptosis is a critical process in the www.aging-us.com growth, maturation, and creation of the nervous system [3]. Many studies have shown that neuronal apoptosis is linked to neural tube deformation in children. Apoptosis happens at a precise time and location during normal embryonic development, according to developmental biology studies. If this carefully controlled, planned cell death is disrupted, it can result in a variety of congenital abnormalities [4]

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