Preclinical Application of Reduced Manipulated Processing Strategy to Collect Transplantable Hepatocytes: A Pilot and Feasibility Study

Ya-Hui Chen; Hui-Ling Chen; Cheng-Maw Ho; Hung-Yen Chen; Shu-Li Ho; Rey-Heng Hu; Po-Huang Lee; Mei-Hwei Chang

doi:10.3390/jpm11050326

Preclinical Application of Reduced Manipulated Processing Strategy to Collect Transplantable Hepatocytes: A Pilot and Feasibility Study

J Pers Med. 2021 Apr 21;11(5):326. doi: 10.3390/jpm11050326.

Authors

Ya-Hui Chen^{1

2}, Hui-Ling Chen¹, Cheng-Maw Ho^{1

3}, Hung-Yen Chen^{1

3}, Shu-Li Ho⁴, Rey-Heng Hu³, Po-Huang Lee³, Mei-Hwei Chang²

Affiliations

¹ Hepatitis Research Center, National Taiwan University Hospital, Taipei 100, Taiwan.
² Department of Pediatrics, National Taiwan University Children Hospital, Taipei 100, Taiwan.
³ Department of Surgery, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei 100, Taiwan.
⁴ Department of Anatomy and Cell Biology, National Yang Ming Chiao Tung University, Taipei 112, Taiwan.

Abstract

Background: The complex isolation and purification process of hepatocytes for transplantation is labor intensive and with great contamination risk. Here, as a pilot and feasibility study, we examined in vitro and in vivo hepatocyte isolation feasibility and cell function of Cell Saver^® Elite^®, an intraoperative blood-cell-recovery system.

Methods: Rat and pig liver cells were collected using this system and then cultured in vitro, and their hepatocyte-specific enzymes were characterized. We then transplanted the hepatocytes in an established acute liver-injured (retrorsine+D-galactosamine-treated) rat model for engraftment. Recipient rats were sacrificed 1, 2, and 4 weeks after transplantation, followed by donor-cell identification and histological, serologic, and immunohistopathological examination. To demonstrate this Cell Saver^® strategy is workable in the first place, traditional (classical) strategy, in our study, behaved as certainty during the cell manufacturing process for monitoring quality assurance throughout the course, from the start of cell isolation to post-transplantation.

Results: We noted that in situ collagenase perfusion was followed by filtration, centrifugation, and collection in the Cell Saver^® until the process ended. Most (>85%) isolated cells were hepatocytes (>80% viability) freshly demonstrating hepatocyte nuclear factor 4α and carbamoyl-phosphate synthase 1 (a key enzyme in the urea cycle), and proliferating through intercellular contact in culture, with expression of albumin and CYP3A4. After hepatocyte transplantation in dipeptidyl peptidase IV (-/-) rat liver, wild-type donor hepatocytes engrafted and repopulated progressively in 4 weeks with liver functional improvement. Proliferating donor hepatocyte-native biliary ductular cell interaction was identified. Post-transplantation global liver functional recovery after Cell Saver and traditional methods was comparable.

Conclusions: Cell Saver^® requires reduced manual manipulation for isolating transplantable hepatocytes.

Keywords: cell processing; cell therapy; hepatocyte; reduced manipulated strategy.

Abstract

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