Long-term and non-invasive in vivo tracking of DiD dye-labeled human hepatic progenitors in chronic liver disease models

Chaturvedula Tripura; Srinivas Gunda; Sandeep Kumar Vishwakarma; Avinash Raj Thatipalli; Jedy Jose; Mahesh Kumar Jerald; Aleem Ahmed Khan; Gopal Pande

doi:10.4254/wjh.v14.i10.1884

Long-term and non-invasive in vivo tracking of DiD dye-labeled human hepatic progenitors in chronic liver disease models

World J Hepatol. 2022 Oct 27;14(10):1884-1898. doi: 10.4254/wjh.v14.i10.1884.

Authors

Chaturvedula Tripura¹, Srinivas Gunda², Sandeep Kumar Vishwakarma³, Avinash Raj Thatipalli², Jedy Jose², Mahesh Kumar Jerald², Aleem Ahmed Khan³, Gopal Pande²

Affiliations

¹ Cell and Stem Cell Biology, CSIR-Centre for Cellular and Molecular Biology, Hyderabad 500007, Telangana, India. tripura@ccmb.res.in.
² Cell and Stem Cell Biology, CSIR-Centre for Cellular and Molecular Biology, Hyderabad 500007, Telangana, India.
³ Central Laboratory for Stem Cell Research and Translational Medicine, Centre for Liver Research and Diagnostics, Deccan College of Medical Sciences, Hyderabad 500058, Telangana, India.

Abstract

Background: Chronic liver diseases (CLD) are the major public health burden due to the continuous increasing rate of global morbidity and mortality. The inherent limitations of organ transplantation have led to the development of stem cell-based therapy as a supportive and promising therapeutic option. However, identifying the fate of transplanted cells in vivo represents a crucial obstacle.

Aim: To evaluate the potential applicability of DiD dye as a cell labeling agent for long-term, and non-invasive in vivo tracking of transplanted cells in the liver.

Methods: Magnetically sorted, epithelial cell adhesion molecule positive (1 × 10⁶ cells/mL) fetal hepatic progenitor cells were labeled with DiD dye and transplanted into the livers of CLD-severe combined immunodeficiency (SCID) mice. Near-infrared (NIR) imaging was performed for in vivo tracking of the DiD-labeled transplanted cells along with colocalization of hepatic markers for up to 80 d. The existence of human cells within mouse livers was identified using Alu polymerase chain reaction and sequencing.

Results: NIR fluorescence imaging of CLD-SCID mice showed a positive fluorescence signal of DiD at days 7, 15, 30, 45, 60, and 80 post-transplantation. Furthermore, positive staining of cytokeratin, c-Met, and albumin colocalizing with DiD fluorescence clearly demonstrated that the fluorescent signal of hepatic markers emerged from the DiD-labeled transplanted cells. Recovery of liver function was also observed with serum levels of glutamic-oxaloacetic transaminase, glutamate-pyruvate transaminase, and bilirubin. The detection of human-specific Alu sequence from the transplanted mouse livers provided evidence for the survival of transplanted cells at day 80.

Conclusion: DiD-labeling is promising for long-term and non-invasive in vivo cell tracking, and understanding the regenerative mechanisms incurred by the transplanted cells.

Keywords: Cell tracking and imaging; Cell transplantation; Chronic liver diseases; DiD; Hepatic progenitors.