Tracing GFP-labeled WJMSCs in vivo using a chronic salpingitis model: an animal experiment

Stem Cell Res Ther. 2017 Dec 1;8(1):272. doi: 10.1186/s13287-017-0714-z.

Abstract

Background: The present study was conducted to evaluate the distribution of Wharton's jelly-derived mesenchymal stem cells (WJMSCs) and their repairing function on the oviduct.

Methods: WJMSCs were transfected with the LV3-GFP-PURO lentivirus. Female New Zealand rabbits (n = 24) were divided randomly into control A and B groups and experimental C and D groups to establish inflammation models. Sterile saline solution or WJMSCs were injected into rabbits via ear veins and/or genital tract perfusion once weekly for 3 weeks. All rabbits were humanely sacrificed 1 week after the last perfusion to collect the oviduct, uterus, liver, and bladder for examination. Green fluorescent protein (GFP) and cytokeratin 7 (CK7) were imaged using a Leica Qwin Plus V3 fluorescence confocal microscope and analyzed as mean optical densities in an Image-Pro Plus analysis system.

Results: We found that lentivirus expressing the GFP gene produced an efficient transfection. The mean optical density values of GFP and CK7 in the oviducts were higher in the experimental D group than those in the control A and experimental C groups. No GFP fluorescence deposits occurred in the bladder of the control A group or experimental C group. Colocalization of CK7 and WJMSCs was observed in the oviducts in all groups.

Conclusions: WJMSCs exhibited homing characteristics and migrated to the injured oviduct to promote epithelial cell growth. Additionally, local treatment resulted in higher efficiency.

Keywords: Green fluorescent protein; Oviducts; Tracing; Transformation; Wharton’s jelly-derived mesenchymal stem cells.

MeSH terms

  • Animals
  • Cell Differentiation
  • Cell Tracking / methods*
  • Disease Models, Animal
  • Epithelial Cells / cytology
  • Epithelial Cells / metabolism
  • Escherichia coli
  • Escherichia coli Infections / microbiology
  • Escherichia coli Infections / pathology
  • Escherichia coli Infections / therapy*
  • Fallopian Tubes / microbiology
  • Fallopian Tubes / pathology
  • Female
  • Gene Expression
  • Genes, Reporter
  • Green Fluorescent Proteins / genetics*
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Keratin-7 / genetics
  • Keratin-7 / metabolism
  • Lentivirus / genetics
  • Lentivirus / metabolism
  • Mesenchymal Stem Cell Transplantation*
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / metabolism
  • Primary Cell Culture
  • Rabbits
  • Salpingitis / microbiology
  • Salpingitis / pathology
  • Salpingitis / therapy*
  • Transfection
  • Transplantation, Heterologous
  • Umbilical Cord / cytology
  • Umbilical Cord / metabolism
  • Wharton Jelly / cytology
  • Wharton Jelly / metabolism

Substances

  • Keratin-7
  • Green Fluorescent Proteins