Abstract
The pig is an important model organism for biomedical research, mainly due to its extensive genetic, physiological and anatomical similarities with humans. Until date, direct conversion of somatic cells into hepatocyte-like cells (iHeps) has only been achieved in rodents and human cells. Here, we employed lentiviral vectors to screen a panel of 12 hepatic transcription factors (TF) for their potential to convert porcine fibroblasts into hepatocyte-like cells. We demonstrate for the first time, hepatic conversion of porcine somatic cells by over-expression of CEBPα, FOXA1 and HNF4α2 (3TF-piHeps). Reprogrammed 3TF-piHeps display a hepatocyte-like morphology and show functional characteristics of hepatic cells, including albumin secretion, Dil-AcLDL uptake, storage of lipids and glycogen and activity of cytochrome P450 enzymes CYP1A2 and CYP2C33 (CYP2C9 in humans). Moreover, we show that markers of mature hepatocytes are highly expressed in 3TF-piHeps, while fibroblastic markers are reduced. We envision piHeps as useful cell sources for future studies on drug metabolism and toxicity as well as in vitro models for investigation of pig-to-human infectious diseases.
Highlights
The pig is an important model organism for biomedical research, mainly due to its extensive genetic, physiological and anatomical similarities with humans
porcine kidney fibroblasts (PKFs) transduced at MOI of 5 per transcription factors (TF)-encoding lentiviral vector showed a significantly higher albumin secretion compared to other MOIs (Fig. 1B), whereas total gene expression levels of hepatic markers such as ALBUMIN, Alpha-1-antitrypsin—A1AT; Transthyretin—TTR,Transferrin—TF and Alpha-fetoprotein—AFP, were induced in cells transduced with 12 TFs, regardless of MOI (Fig. 1C)
Cellular reprogramming by over-expression of exogenous factors was first described in 1987, when mouse embryonic fibroblasts were converted into myoblasts by over-expression of MyoD31
Summary
The pig is an important model organism for biomedical research, mainly due to its extensive genetic, physiological and anatomical similarities with humans. Pigs have a long standing and very successful history as biomedical model for studying human diseases and developing novel therapies, which is mainly attributed to the many genetic, anatomical and physiological similarities with humans[1,2,3]. This resemblance renders pigs important models for developing novel surgical techniques[4], endoscopic approaches, such as NOTES (natural orifice transluminal endoscopic surgery)[5] and even for complex metabolic disorders[6]. Hepatocyte-like cells could provide a more stable and renewable cell source with on-demand availability for scientific and therapeutic studies on those important topics
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