P190\u2005Proof of concept & novel technique of cell harvest using histotripsy: implications in cell transplantation

Abstract

Introduction A potential alternative to liver transplantation is allogenic hepatocyte transplantation, particularly for metabolic disorders. However, some significant hurdles mainly concerned shortage of donor organs, low cell yield as well as lack of long-standing effect needs to be overcome to widen its clinical application. Here we describe an improved technique in cell harvest and isolation. Methods Pig livers were obtained using organ retrieval techniques and perfused with Soltran solution following a period of cold storage. Perfused livers were subjected to High-Intensity Focused Ultrasound (HIFU), and lesions were incised. Core liquified suspension was sampled and cultured in RPMI cell culture medium. Cell cultures were analysed at 1, 3 and 7 days for viability. H&E staining performed to characterise the lesions. Results Four different livers were used, and more than 50 lesions created. HIFU created a subcapsular lesion with a core suspension of cells. Adult hepatocytes extracted from core lesion are alive at day 1 and remain alive in culture medium for up to 1 weeks. Live-dead assay confirms cell viability with retainment of metabolic activity and morphology at 1 week. Cells also have increased in number from 0.12% live cells on day 1 to 0.45% on day 7 (P Discussion HIFU has been used in treating benign and cancerous lesion has shown promising results. However, a particular modality of HIFU, boiling histotripsy, can be used to increase the yield of adult hepatocytes extraction & isolation safely. Here we report a detergent and chemical-free cell harvest technique. It can improve the quality and number of cells for transplantation. Further studies are required to asses long term effect extracted cells.