Journal Article

Tbx3 improves the germ-line competency of induced pluripotent stem cells

Tay, Junliang, Yuan, Ping, Soh, Boon Seng, Li, Pin, Orlov, Yuriy L., Han, Jianyong, Yang, Henry, Lim, Bing, Cao, Suying, Lim, Siew Lan, Lufkin, Thomas, Zhang, Jinqiu, Ng, Huck-Hui, Tam, Wai-Leong

2010

Overview

Tbx3 boosts iPS quality While much attention has been given to the study of different genetic and chemical methods for the generation of iPS (induced pluripotent stem) cell lines, relatively little is known about the variability in overall quality of iPS cells. This paper identifies a transcription factor, Tbx3, that significantly improves the quality of iPS cells. Tbx3 also accelerates the reprogramming process of mouse embryonic fibroblasts into iPS cells and significantly improves the germ-line transmission of iPS-derived germ cells in chimaeric animals. The transcription factor Tbx3 is shown to significantly improve the quality of induced pluripotent stem (iPS) cells. Tbx3 binding sites in embryonic stem cells are present in genes involved in pluripotency and reprogramming factors. Furthermore, there are intrinsic qualitative differences in iPS cells generated by different methods in terms of their pluripotency, thus highlighting the need to rigorously characterize iPS cells beyond in vitro studies. Induced pluripotent stem (iPS) cells can be obtained by the introduction of defined factors into somatic cells 1 . The combination of Oct4 (also known as Pou5f1), Sox2 and Klf4 (which we term OSK) constitutes the minimal requirement for generating iPS cells from mouse embryonic fibroblasts. These cells are thought to resemble embryonic stem cells (ESCs) on the basis of global gene expression analyses; however, few studies have tested the ability and efficiency of iPS cells to contribute to chimaerism, colonization of germ tissues, and most importantly, germ-line transmission and live birth from iPS cells produced by tetraploid complementation. Using genomic analyses of ESC genes that have roles in pluripotency and fusion-mediated somatic cell reprogramming, here we show that the transcription factor Tbx3 significantly improves the quality of iPS cells. iPS cells generated with OSK and Tbx3 (OSKT) are superior in both germ-cell contribution to the gonads and germ-line transmission frequency. However, global gene expression profiling could not distinguish between OSK and OSKT iPS cells. Genome-wide chromatin immunoprecipitation sequencing analysis of Tbx3-binding sites in ESCs suggests that Tbx3 regulates pluripotency-associated and reprogramming factors, in addition to sharing many common downstream regulatory targets with Oct4, Sox2, Nanog and Smad1. This study underscores the intrinsic qualitative differences between iPS cells generated by different methods, and highlights the need to rigorously characterize iPS cells beyond in vitro studies.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 631/136/532/2064/2158 / 631/45/612/822 / 631/532/2435 / Analysis / Animals / Biological and medical sciences / Cell differentiation / Cell Fusion / Cellular Reprogramming / Chemical compounds / Chimera - embryology / Chimera - metabolism / Chromatin Immunoprecipitation / Drug resistance / Efficiency / Embryo, Mammalian - cytology / Embryonic stem cells / Female / Fibroblasts - cytology / Fibroblasts - metabolism / Fundamental and applied biological sciences. Psychology / Gene Expression Profiling / Gene Expression Regulation - genetics / Genetic aspects / Germ Cells - cytology / Germ Cells - metabolism / Gonads / Gonads - cytology / Homeodomain Proteins - metabolism / Humanities and Social Sciences / Induced Pluripotent Stem Cells - cytology / Induced Pluripotent Stem Cells - metabolism / Kinases / Kruppel-Like Factor 4 / Kruppel-Like Transcription Factors - genetics / Kruppel-Like Transcription Factors - metabolism / letter / Male / Mice / Mice, Transgenic / Molecular and cellular biology / multidisciplinary / Nanog Homeobox Protein / Octamer Transcription Factor-3 - genetics / Octamer Transcription Factor-3 - metabolism / Physiological aspects / Proteins / Regulatory Sequences, Nucleic Acid / Science / Science (multidisciplinary) / Smad1 Protein - metabolism / SOXB1 Transcription Factors - genetics / SOXB1 Transcription Factors - metabolism / Stem cells / T-Box Domain Proteins - genetics / T-Box Domain Proteins - metabolism / Transcription factors / Transcription, Genetic - genetics / Transduction, Genetic