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

Zhe Li,Zhao Zhang,Wai-Kit Ming,Xin Chen,Xiao-Min Xiao

doi:10.1186/s13287-017-0714-z

Abstract

BackgroundThe present study was conducted to evaluate the distribution of Wharton’s jelly-derived mesenchymal stem cells (WJMSCs) and their repairing function on the oviduct.MethodsWJMSCs 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.ResultsWe 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.ConclusionsWJMSCs exhibited homing characteristics and migrated to the injured oviduct to promote epithelial cell growth. Additionally, local treatment resulted in higher efficiency.

Highlights

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
Assessment of transfection efficiency of LV3-Green fluorescent protein (GFP)-Puro lentivirus in vitro Cells were transduced with LV3-GFP-PURO lentivirus (5 × 109 TU/ml) at 100, 200, and 400 MOI for 96 h, and all WJMSCs expressed green fluorescence
Optimal transfection efficiency of LV3-GFPPuro lentivirus occurred at 200 MOI, and 200 MOI was used in subsequent experiments (Fig. 1)

Summary

Introduction

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. 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. Salpingitis and/or pelvic inflammation caused by infection is one of the most important factors of tubal infertility. New methods to repair the reproduction function of the oviduct are required to allow gametes to combine and grow in vivo under natural circumstances. Mesenchymal stem cells (MSCs) [2] are used in cell therapy and regenerative medicine because these cells are our study established a chronic salpingitis model in New Zealand rabbits and transplanted WJMSCs expressing green fluorescent protein (GFP)

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