Research Paper Volume 13, Issue 17 pp 21778—21790
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
- 1 Department of Pediatrics, The First Affiliated Hospital of Harbin Medical University, Harbin 150001, P.R. China
- 2 Department of Ultrasound Medicine, The Fifth Hospital of Harbin, Harbin 150040, P.R. China
- 3 Department of Neurosurgery, Tieling Central Hospital, Tieling 112000, P.R. China
- 4 Department of Stomatology, The Fourth Affiliated Hospital of Harbin Medical University, Harbin 150001, P.R. China
- 5 Department of Pediatrics, The First Hospital of Jilin University, Changchun 130021, P.R. China
Received: January 18, 2021 Accepted: September 3, 2021 Published: September 14, 2021
https://doi.org/10.18632/aging.203524How to Cite
Copyright: © 2021 Jiang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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.
Abbreviations
STAT3: signal transducer and activator of transcription-3; BMSCs: bone mesenchymal Stem Cells; SBA: spina bifida aperta; E20: 20-day pregnant; NTDs: neural tube defects; GFP: green fluorescent protein; RA: retinoic acid; NF: neuromodulatory cytokine; OD: optical density; CCK-8: Cell Counting Kit-8; LIF: leukemia inhibitory factor; CNTF: ciliary neurotrophic factor; GFAP: glial fibrillary acidic protein; TNFR1: tumor necrosis factor receptor type 1; TRAIL: TNF-related apoptosis-inducing ligand; MAPK: mitogen-activated protein kinase.