Expansion of functional personalized cells with specific transgene combinations

Christoph Lipps,Joachim R Göthert,Jörn Pezoldt,Jochen Hühn,Mario Köster,Hansjörg Hauser,Carolina Sañudo,Michael Ott,Virginia Seiffert,Tom Wahlicht,Wolf Drescher,Milada Butueva,Martin Luckner,Oliver Dittrich‐Breiholz ,Qing Yuan ,Peter Müller ,Rainer Beckmann,Henning Kempf,Paula M Alves,Roland Schucht,Dagmar Wirth,Vera Rebmann,Theresa Truschel,Thomas Pufe,Jeannette Zauers,Thomas Korff,Friederike Klein ,José A Riancho ,Roderick A.f Macleod ,Robert Zweigerdt,Bertram Opalka,Tobias May

doi:10.1038/s41467-018-03408-4

Abstract

Fundamental research and drug development for personalized medicine necessitates cell cultures from defined genetic backgrounds. However, providing sufficient numbers of authentic cells from individuals poses a challenge. Here, we present a new strategy for rapid cell expansion that overcomes current limitations. Using a small gene library, we expanded primary cells from different tissues, donors, and species. Cell-type-specific regimens that allow the reproducible creation of cell lines were identified. In depth characterization of a series of endothelial and hepatocytic cell lines confirmed phenotypic stability and functionality. Applying this technology enables rapid, efficient, and reliable production of unlimited numbers of personalized cells. As such, these cell systems support mechanistic studies, epidemiological research, and tailored drug development.

Highlights

Fundamental research and drug development for personalized medicine necessitates cell cultures from defined genetic backgrounds
Long-term expandable cell cultures were established from all tested cell types including chondrocytes, epithelial, and endothelial cells and hepatocytes showing the wide applicability of the library for immortalization of diverse cell types
When we analyzed e-hUVEC-2 cells for nitric oxide (NO) production and acetylated low-density lipoprotein uptake, we found comparable levels after 45 and 90 cumulative population doublings demonstrating a stable phenotype upon cultivation (Fig. 3d)

Summary

Introduction

Fundamental research and drug development for personalized medicine necessitates cell cultures from defined genetic backgrounds. In depth characterization of a series of endothelial and hepatocytic cell lines confirmed phenotypic stability and functionality Applying this technology enables rapid, efficient, and reliable production of unlimited numbers of personalized cells. As such, these cell systems support mechanistic studies, epidemiological research, and tailored drug development. Cardiology, Justus-Liebig University Giessen, Aulweg 129, 35392 Giessen, Germany These authors contributed : Christoph Lipps, Franziska Klein, Tom. Cell culture is an essential tool to study the fundamentals of genetic background variables. The cellular gene encoding human telomerase reverse transcriptase (hTert) has been successfully used for the expansion of several cell types that retain important properties in vitro[13,14,15]. This approach is restricted to certain cell types as others either need the concerted action of additional immortalizing genes or the inactivation of tumor suppressor genes[16,17,18,19]

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Conclusion