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

Aging (Albany NY). 2021 Sep 14;13(17):21778-21790. doi: 10.18632/aging.203524. Epub 2021 Sep 14.

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.

Keywords: bone marrow mesenchymal stem cells; lentivirus transfection; nerve cells; signal transducer and activator of transcription-3; spina bifida aperta.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Marrow Cells / physiology
  • Cell Differentiation
  • Female
  • Fetus / metabolism
  • Fetus / surgery*
  • Lentivirus / genetics
  • Lentivirus / metabolism
  • Male
  • Mesenchymal Stem Cell Transplantation*
  • Mesenchymal Stem Cells / metabolism*
  • Nestin
  • Plasmids
  • Rats
  • Rats, Wistar
  • STAT3 Transcription Factor / metabolism*
  • Spina Bifida Cystica / metabolism
  • Spina Bifida Cystica / therapy*
  • Spinal Cord / metabolism
  • Tissue Engineering*
  • Transfection
  • Tretinoin

Substances

  • NES protein, human
  • Nestin
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • Tretinoin