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Compensatory proliferation of endogenous chicken primordial germ cells after elimination by busulfan treatment

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Authors
Lee, Hyung Chul; Kim, Sung Kyu; Park, Tae Sub; Rengaraj, Deivendran; Park, Kyung Je; Lee, Hong Jo; Park, Soo Bong; Kim, Sung Woo; Choi, Seong Bok; Han, Jae Yong
Issue Date
2013
Publisher
BioMed Central
Citation
Stem Cell Research and Therapy, 4:136
Description
This is an open access article distributed under the terms of the Creative
Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Introduction
Primordial germ cells (PGCs) are the major population of cells in the developing bilateral embryonic gonads. Little is known about the cellular responses of PGCs after treatment with toxic chemicals such as busulfan during embryo development. In this study, we investigated the elimination, restorative ability, and cell cycle status of endogenous chicken PGCs after busulfan treatment.

Methods
Busulfan was emulsified in sesame oil by a dispersion-emulsifying system and injected into the chick blastoderm (embryonic stage X). Subsequently, we conducted flow cytometry analysis to evaluate changes in the PGC population and cell cycle status, and immunohistochemistry to examine the germ cell proliferation.

Results
Results of flow cytometry and immunohistochemistry analyses after busulfan treatment showed that the proportion of male PGCs at embryonic day 9 and female PGCs at embryonic day 7 were increased by approximately 60% when compared with embryonic day 5.5. This result suggests the existence of a compensatory mechanism in PGCs in response to the cytotoxic effects of busulfan. Results of cell cycling analysis showed that the germ cells in the G0/G1 phase were significantly decreased, while S/G2/M-phase germ cells were significantly increased in the treatment group compared with the untreated control group in both 9-day-old male and female embryos. In addition, in the proliferation analysis with 5-ethynyl-2′-deoxyuridine (EdU) incorporation, we found that the proportion of EdU-positive cells among VASA homolog-positive cells in the 9-day embryonic gonads of the busulfan-treated group was significantly higher than in the control group.

Conclusions
We conclude that PGCs enter a restoration pathway by promoting their cell cycle after experiencing a cytotoxic effect.
ISSN
1757-6512
Language
English
URI
http://stemcellres.com/content/4/6/136

https://hdl.handle.net/10371/100371
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College of Agriculture and Life Sciences (농업생명과학대학)Dept. of Food and Animal Biotechnology (식품·동물생명공학부)Journal Papers (저널논문_식품·동물생명공학부)
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