Abstract

Determining the cell fate and the distribution of mesenchymal stromal/stem cells (MSCs) after transplantation are essential parts of characterizing the mechanisms of action and biosafety profile of stem cell therapy. Many recent studies have shown that MSCs migrate into injured tissues, but are only detectable at extremely low frequencies. We investigated the cell fate of MSCs after transplantation in an acute kidney injury (AKI) mouse model using in vivo bioluminescence imaging (BLI) and subsequent verification of cell migration using quantitative real-time polymerase chain reaction (qRT-PCR). The AKI was induced by a single injection of cisplatin (8 or 12 mg/kg). One day later, adipose-derived mesenchymal stromal/stem cells isolated from luciferase transgenic mice (Luc+-mASCs, 5 × 105) were intravenously transplanted. Migration kinetics of the cells was monitored using BLI on day 1, 3, and 6, and finally via quantitative real-time PCR at the endpoint on day 6. Using BLI, infused Luc+-mASCs could only be detected in the lungs, but not in the kidneys. In contrast, PCR endpoint analysis revealed that Luc-specific mRNA could be detected in injured renal tissue; compared to the control group, the induction was 2.2-fold higher for the 8 mg/kg cisplatin group (p < 0.05), respectively 6.1-fold for the 12 mg/kg cisplatin group (p < 0.001). In conclusion, our study demonstrated that Luc-based real-time PCR rather than BLI is likely to be a better tool for cell tracking after transplantation in models such as cisplatin-induced AKI.

Highlights

  • Determining the cell fate and the distribution of mesenchymal stromal/stem cells (MSCs) after transplantation are essential parts of characterizing the mechanisms of action and biosafety profile of stem cell therapy [1]

  • Our study demonstrated that Luc-based real-time PCR rather than bioluminescence imaging (BLI) is likely to be a better tool for cell tracking after transplantation in models such as cisplatin-induced acute kidney injury (AKI)

  • We investigated the cell fate of MSCs after transplantation in an AKI model using in vivo BLI and subsequent verification of luciferase mRNA in the kidney using quantitative real-time reverse transcriptase-PCR

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Summary

Introduction

Determining the cell fate and the distribution of mesenchymal stromal/stem cells (MSCs) after transplantation are essential parts of characterizing the mechanisms of action and biosafety profile of stem cell therapy [1]. The therapeutic use of transplanted MSCs by intravenous (IV) administration was investigated by in vivo studies and injury models These in vivo studies used various techniques to label and track culture-expanded and transplanted MSCs in different tissues over time, e.g., cell labeling by fluorescence or radioactivity, transfection with reporter genes (luciferase or green fluorescent protein), or the use of donor cell-specific markers (human Alu sequence) [1,2,3,4,5,6,7]. We investigated the cell fate of MSCs after transplantation in an AKI model using in vivo BLI and subsequent verification of luciferase mRNA in the kidney using quantitative real-time reverse transcriptase-PCR (qRT-PCR)

Characterization of mASC
In Vitro Bioluminescence and PCR Analysis
In Vivo Biolumunescence Imaging
Animals
In Vivo Model of Cisplatin-Induced Acute Kidney Injury
Polymerase Chain Reaction
In Vivo Bioluminescence Imaging
Statistical Analysis
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