Stably transduction of J2-3T3 fibroblasts using lentivirus and ViroMag R/L

J2-3T3 fibroblasts were generated by ViroMag R/L mediated transduction of lentivirus and subsequent selection with puromycin.

This paper shows the efficiency of ViroMag R/L from OZ Biosciences to transduce with high efficiency J2-3T3 fibroblasts for subsequent selectio.

article reference: Tissue Eng Part A. 2014 Feb 5. 

Micropatterned cell-cell interactions enable functional encapsulation of primary hepatocytes in hydrogel microtissues.

Li CY, Stevens KR, Schwartz RE, Alejandro BS, Huang JH, Bhatia SN.

Abstract

Drug-induced liver injury is a major cause of drug development failures and post-market withdrawals. In vitro models that incorporate primary hepatocytes have been shown to be more predictive than model systems that rely on liver microsomes or hepatocellular carcinoma cell lines. Methods to phenotypically stabilize primary hepatocytes ex vivo often rely on mimicry of hepatic microenvironmental cues such as cell-cell interactions and cell-matrix interactions. In this work, we sought to incorporate phenotypically-stable hepatocytes into 3D microtissues, which in turn could be deployed in drug screening platforms such as multiwell plates and diverse organ-on-a-chip devices. We first utilize micropatterning on collagen I to specify cell-cell interactions in 2D, followed by collagenase digestion to produce well-controlled aggregates for 3D encapsulation in PEG diacrylate. Using this approach, we examined the influence of homotypic hepatocyte interactions and composition of the encapsulating hydrogel, and achieved maintenance of liver-specific function for over 50 days. Optimally pre-aggregated structures were subsequently encapsulated using a microfluidic droplet-generator to produce 3D microtissues. Interactions of engineered hepatic microtissues with drugs was characterized by flow cytometry, and yield both induction of P450 enzymes in response to prototypic small molecules, as well as drug-drug interactions that give rise to hepatotoxicity. Collectively, this study establishes a pipeline for the manufacturing of 3D hepatic microtissues that exhibit stabilized liver-specific functions and can be incorporated into a wide array of emerging drug development platforms.

ViroMag R/L from OZ Biosciences is a magnetic nanoparticles formulation optimized for increasing retro- and lentivirus infection and transduction both in vitro and in vivo.