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Mol. Cells 2010; 29(5): 519-526

Published online May 31, 2010

https://doi.org/10.1007/s10059-010-0064-5

© The Korean Society for Molecular and Cellular Biology

Identification of an Intermediate State as Spermatogonial Stem Cells Reprogram to Multipotent Cells

Hyung Joon Kim1,2, Hyun Jung Lee1, Jung Jin Lim1,2, Ki Hoon Kwak1, Jong Soo Kim2, Ji Hoon Kim3, Yong-Mahn Han3, Kye-Seong Kim2,*, and Dong Ryul Lee1,4,*

1Fertility Center of CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul 135-081, Korea, 2Department of Anatomy and Cell Biology, College of Medicine, Hanyang University, Seoul 133-791, Korea, 3Department of Biological Sciences and Center for Stem Cell Differentiation, Korea Advanced Institute of Science and Technology, Daejeon 305-701, Korea, 4Department of Biomedical Science, College of Life Science, CHA University, Seoul 135-081, Korea

Correspondence to : *Correspondence: drleedr@cha.ac.kr (DRL); ks66kim@hanyang.ac.kr (KK)

Received: December 23, 2010; Accepted: January 7, 2010

Abstract

The aim of this study was to understand the mechanisms that allow mSSC lines to be established from SSCs. Small, multilayer clumps of SSCs formed during two to four weeks of in vitro culture and were then transferred to MEF feeders. Small, round, monolayer colonies containing cells destined to convert to mSSCs, designated as intermediate state SSCs (iSSCs), first appeared after two to three passages. During an additional nine passages (47-54 days) under the same culture conditions, iSSCs slowly proliferated and maintained their morphology. Ultimately, a cell type with an ES-like morphology (mSSC) appeared from the iSSC colonies, and two mSSC cell lines were established. The mSSCs had a high proliferative potential in serum-free ES culture medium and have been successfully maintained since their first establishment (> 12 months). We also compared the specific characteristics of iSSCs with those of SSCs and mSSCs using immunocytochemistry, FACS, RT-PCR, DNA methylation, and miRNA analyses. The results suggest that iSSCs represent a morpholog-ically distinct intermediate state with characteristic ex-pression patterns of pluripotency-related genes and miR-NAs that arise during the conversion of SSCs into mSSCs. Our results suggest that iSSCs could be a useful model for evaluating and understanding the initiation mechanisms of cell reprogramming.

Keywords culture-induced reprogramming, dedifferentiation, intermediate state, miRNA, spermatogonial stem cells

Article

Research Article

Mol. Cells 2010; 29(5): 519-526

Published online May 31, 2010 https://doi.org/10.1007/s10059-010-0064-5

Copyright © The Korean Society for Molecular and Cellular Biology.

Identification of an Intermediate State as Spermatogonial Stem Cells Reprogram to Multipotent Cells

Hyung Joon Kim1,2, Hyun Jung Lee1, Jung Jin Lim1,2, Ki Hoon Kwak1, Jong Soo Kim2, Ji Hoon Kim3, Yong-Mahn Han3, Kye-Seong Kim2,*, and Dong Ryul Lee1,4,*

1Fertility Center of CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul 135-081, Korea, 2Department of Anatomy and Cell Biology, College of Medicine, Hanyang University, Seoul 133-791, Korea, 3Department of Biological Sciences and Center for Stem Cell Differentiation, Korea Advanced Institute of Science and Technology, Daejeon 305-701, Korea, 4Department of Biomedical Science, College of Life Science, CHA University, Seoul 135-081, Korea

Correspondence to:*Correspondence: drleedr@cha.ac.kr (DRL); ks66kim@hanyang.ac.kr (KK)

Received: December 23, 2010; Accepted: January 7, 2010

Abstract

The aim of this study was to understand the mechanisms that allow mSSC lines to be established from SSCs. Small, multilayer clumps of SSCs formed during two to four weeks of in vitro culture and were then transferred to MEF feeders. Small, round, monolayer colonies containing cells destined to convert to mSSCs, designated as intermediate state SSCs (iSSCs), first appeared after two to three passages. During an additional nine passages (47-54 days) under the same culture conditions, iSSCs slowly proliferated and maintained their morphology. Ultimately, a cell type with an ES-like morphology (mSSC) appeared from the iSSC colonies, and two mSSC cell lines were established. The mSSCs had a high proliferative potential in serum-free ES culture medium and have been successfully maintained since their first establishment (> 12 months). We also compared the specific characteristics of iSSCs with those of SSCs and mSSCs using immunocytochemistry, FACS, RT-PCR, DNA methylation, and miRNA analyses. The results suggest that iSSCs represent a morpholog-ically distinct intermediate state with characteristic ex-pression patterns of pluripotency-related genes and miR-NAs that arise during the conversion of SSCs into mSSCs. Our results suggest that iSSCs could be a useful model for evaluating and understanding the initiation mechanisms of cell reprogramming.

Keywords: culture-induced reprogramming, dedifferentiation, intermediate state, miRNA, spermatogonial stem cells

Mol. Cells
Nov 30, 2022 Vol.45 No.11, pp. 763~867
COVER PICTURE
Naive (cyan) and axotomized (magenta) retinal ganglion cell axons in Xenopus tropicalis (Choi et al., pp. 846-854).

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