Metastable Reprogramming State of Single Transcription Factor-Derived Induced Hepatocyte-Like Cells

We previously described the generation of induced hepatocyte-like cells (iHeps) using the hepatic transcription factor Hnf1a together with small molecules. These iHeps represent a hepatic state that is more mature compared with iHeps generated with multiple hepatic factors. However, the underlying m...

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Main Authors: Seon In Hwang, Tae Hwan Kwak, Ji Hyun Kang, Jonghun Kim, Hyunseong Lee, Kee-Pyo Kim, Kinarm Ko, Hans R. Schöler, Dong Wook Han
Format: Article
Language:English
Published: Wiley 2019-01-01
Series:Stem Cells International
Online Access:http://dx.doi.org/10.1155/2019/6937257
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author Seon In Hwang
Tae Hwan Kwak
Ji Hyun Kang
Jonghun Kim
Hyunseong Lee
Kee-Pyo Kim
Kinarm Ko
Hans R. Schöler
Dong Wook Han
author_facet Seon In Hwang
Tae Hwan Kwak
Ji Hyun Kang
Jonghun Kim
Hyunseong Lee
Kee-Pyo Kim
Kinarm Ko
Hans R. Schöler
Dong Wook Han
author_sort Seon In Hwang
collection DOAJ
description We previously described the generation of induced hepatocyte-like cells (iHeps) using the hepatic transcription factor Hnf1a together with small molecules. These iHeps represent a hepatic state that is more mature compared with iHeps generated with multiple hepatic factors. However, the underlying mechanism of hepatic conversion involving transgene dependence of the established iHeps is largely unknown. Here, we describe the generation of transgene-independent iHeps by inducing the ectopic expression of Hnf1a using both an episomal vector and a doxycycline-inducible lentivirus. In contrast to iHeps with sustained expression of Hnf1a, transgene-independent Hnf1a iHeps lose their typical morphology and in vitro functionality with rapid downregulation of hepatic markers upon withdrawal of small molecules. Taken together, our data indicates that the reprogramming state of single factor Hnf1a-derived iHeps is metastable and that the hepatic identity of these cells could be maintained only by the continuous supply of either small molecules or the master hepatic factor Hnf1a. Our findings emphasize the importance of a factor screening strategy for inducing specific cellular identities with a stable reprogramming state in order to eventually translate direct conversion technology to the clinic.
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spelling doaj-art-28ac5f0f5ff1484e9d31ae5dd7ccf5c92025-08-20T02:03:54ZengWileyStem Cells International1687-966X1687-96782019-01-01201910.1155/2019/69372576937257Metastable Reprogramming State of Single Transcription Factor-Derived Induced Hepatocyte-Like CellsSeon In Hwang0Tae Hwan Kwak1Ji Hyun Kang2Jonghun Kim3Hyunseong Lee4Kee-Pyo Kim5Kinarm Ko6Hans R. Schöler7Dong Wook Han8Department of Stem Cell Biology, School of Medicine, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of KoreaDepartment of Stem Cell Biology, School of Medicine, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of KoreaDepartment of Stem Cell Biology, School of Medicine, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of KoreaDepartment of Stem Cell Biology, School of Medicine, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of KoreaDepartment of Stem Cell Biology, School of Medicine, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of KoreaDepartment of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstraße 20, 48149 Münster, GermanyDepartment of Stem Cell Biology, School of Medicine, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of KoreaDepartment of Stem Cell Biology, School of Medicine, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of KoreaDepartment of Stem Cell Biology, School of Medicine, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of KoreaWe previously described the generation of induced hepatocyte-like cells (iHeps) using the hepatic transcription factor Hnf1a together with small molecules. These iHeps represent a hepatic state that is more mature compared with iHeps generated with multiple hepatic factors. However, the underlying mechanism of hepatic conversion involving transgene dependence of the established iHeps is largely unknown. Here, we describe the generation of transgene-independent iHeps by inducing the ectopic expression of Hnf1a using both an episomal vector and a doxycycline-inducible lentivirus. In contrast to iHeps with sustained expression of Hnf1a, transgene-independent Hnf1a iHeps lose their typical morphology and in vitro functionality with rapid downregulation of hepatic markers upon withdrawal of small molecules. Taken together, our data indicates that the reprogramming state of single factor Hnf1a-derived iHeps is metastable and that the hepatic identity of these cells could be maintained only by the continuous supply of either small molecules or the master hepatic factor Hnf1a. Our findings emphasize the importance of a factor screening strategy for inducing specific cellular identities with a stable reprogramming state in order to eventually translate direct conversion technology to the clinic.http://dx.doi.org/10.1155/2019/6937257
spellingShingle Seon In Hwang
Tae Hwan Kwak
Ji Hyun Kang
Jonghun Kim
Hyunseong Lee
Kee-Pyo Kim
Kinarm Ko
Hans R. Schöler
Dong Wook Han
Metastable Reprogramming State of Single Transcription Factor-Derived Induced Hepatocyte-Like Cells
Stem Cells International
title Metastable Reprogramming State of Single Transcription Factor-Derived Induced Hepatocyte-Like Cells
title_full Metastable Reprogramming State of Single Transcription Factor-Derived Induced Hepatocyte-Like Cells
title_fullStr Metastable Reprogramming State of Single Transcription Factor-Derived Induced Hepatocyte-Like Cells
title_full_unstemmed Metastable Reprogramming State of Single Transcription Factor-Derived Induced Hepatocyte-Like Cells
title_short Metastable Reprogramming State of Single Transcription Factor-Derived Induced Hepatocyte-Like Cells
title_sort metastable reprogramming state of single transcription factor derived induced hepatocyte like cells
url http://dx.doi.org/10.1155/2019/6937257
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