Basic Study
Copyright ©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Hepatol. Oct 27, 2022; 14(10): 1884-1898
Published online Oct 27, 2022. 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
Chaturvedula Tripura, Srinivas Gunda, Sandeep Kumar Vishwakarma, Avinash Raj Thatipalli, Jedy Jose, Mahesh Kumar Jerald, Aleem Ahmed Khan, Gopal Pande
Chaturvedula Tripura, Srinivas Gunda, Avinash Raj Thatipalli, Jedy Jose, Mahesh Kumar Jerald, Gopal Pande, Cell and Stem Cell Biology, CSIR-Centre for Cellular and Molecular Biology, Hyderabad 500007, Telangana, India
Sandeep Kumar Vishwakarma, Aleem Ahmed Khan, Central Laboratory for Stem Cell Research and Translational Medicine, Centre for Liver Research and Diagnostics, Deccan College of Medical Sciences, Hyderabad 500058, Telangana, India
Author contributions: Khan AA, Pande G, and Tripura C were responsible for the study concept, design, and supervision; Thatipalli AR, Vishwakarma SK, Jose J, and Jerald MK performed the experiments; Tripura C, and Gunda S were responsible for data acquisition and analysis; Tripura C also performed data organization and manuscript writing; Tripura C, Vishwakarma SK, Khan AA, and Pande G performed editing and revision of the manuscript draft.
Supported by Department of Science and Technology (DST), Ministry of Science and Technology, Govt. of India and Indian Council of Medical Research (ICMR), New Delhi, Govt. of India Grants to GP, No. GAP-0220 and No. GAP-0383.
Institutional review board statement: The total fetal liver cells (tFLCs) isolation protocol was approved by the Institutional Ethics Committee (IEC) of Deccan College of Medical Sciences (DCMS), Hyderabad.
Institutional animal care and use committee statement: The study was approved by the Institutional Animal Ethics Committee (Animal trial registration number 20/1999/CPCSEA dated 10/03/1999) of CCMB.
Informed consent statement: Informed consent was obtained prior to sample collection, and cell processing was performed according to the ethical guidelines for the use of human cells.
Conflict-of-interest statement: Dr. Tripura reports grants from the Department of Science and Technology (DST, Ministry of Science and Technology, Govt. of India and Indian Council of Medical Research (ICMR), New Delhi, Govt. of India Grants No. GAP-0220 and No. GAP-0383 to GP during the study period.
Data sharing statement: Data will be shared on request.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
Corresponding author: Chaturvedula Tripura, PhD, Senior Scientist, Cell and Stem Cell Biology, CSIR-Centre for Cellular and Molecular Biology, Uppal Road, Habsiguda, Hyderabad 500007, Telangana, India.
Received: July 26, 2022
Peer-review started: July 26, 2022
First decision: August 18, 2022
Revised: September 2, 2022
Accepted: October 2, 2022
Article in press: October 2, 2022
Published online: October 27, 2022
Research background

Determining the fate of transplanted cells in vivo through long-term cell tracking remains a crucial field of investigation. Long-term live cell tracking in vivo has always been challenging due to the absence of a safe cell-labeling agent.

Research motivation

DiD is a carbocyanine dye having good photochemical properties of strong fluorescence, and stability. Due to the long-range emission of DiD (670 nm), tissue autofluorescence is minimum, permitting the use of other fluorochromes such as fluorescein isothiocyanate, for colocalization studies to evaluate the expression of other essential markers specific to the transplanted cells in the recipient tissue. Moreover, the process for labeling the cells using DiD is easy due to its excellent efficiency for integration and diffusion into the cell membranes. The effects of DiD labeling on in vivo retention of labeled human liver cells remain to be investigated.

Research objectives

The present study aimed to shed light on the fate of DiD-labeled human liver cells in chronic liver diseases (CLD)-severe combined immunodeficiency (SCID) mice using live imaging up to 80 d post-transplantation.

Research methods

A chronic liver disease SCID mouse model was developed, which received DiD-labeled EpCAM-positive human hepatic progenitor cells by intra-hepatic infusion. The long-term survival and functional response of transplanted DiD-labeled cells were investigated up to 80 d.

Research results

This study showed that DiD labeling of human liver cells is easy and efficient for long-term and non-invasive tracking in vivo up to 80 d post-transplantation. Using DiD, the fate of transplanted cells was determined. Transplanted human fetal liver cells were able to provide structural and functional improvement in CLD-SCID mice.

Research conclusions

Monitoring the fate of transplanted cells through DiD-based in vivo live cell imaging can help in understanding the homing, engraftment, long-term survival, and function of the transplanted cells.

Research perspectives

The findings of the current study may pave the way to unravel the underlying regenerative mechanisms and contribution of exogenously transplanted cells in restoring the structural and functional deficits of the liver in CLD.