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Blastocyst Stage Affects the Isolation and Culture of Buffalo Naive/Primed Embryonic Stem-Like Cells

Published online by Cambridge University Press:  13 September 2022

Xin Zhao Open the ORCID record for Xin Zhao [Opens in a new window] ,
Qiuyan Ruan ,
Yun Feng ,
Ziyun Ruan ,
Zhulian Wu ,
Deshun Shi  and
Fenghua Lu
Xin Zhao
Affiliation:
Animal Reproduction Institute, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530005, China Reproductive Medicine Center, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, Nanning 530005, China
Qiuyan Ruan
Affiliation:
Animal Reproduction Institute, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530005, China Research Center for Reproductive Medicine, Reproductive Hospital of Guangxi Zhuang Autonomous Region, Nanning 530005, China
Yun Feng
Affiliation:
Animal Reproduction Institute, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530005, China
Ziyun Ruan
Affiliation:
Animal Reproduction Institute, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530005, China
Zhulian Wu
Affiliation:
Animal Reproduction Institute, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530005, China Reproductive Medicine Center, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, Nanning 530005, China
Deshun Shi*
Affiliation:
Animal Reproduction Institute, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530005, China
Fenghua Lu*
Affiliation:
Animal Reproduction Institute, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530005, China
*
*Corresponding authors: Deshun Shi, Email: ardsshi@gxu.edu.cn, Fenghua Lu, Email: lfhgggg@163.com
*Corresponding authors: Deshun Shi, Email: ardsshi@gxu.edu.cn, Fenghua Lu, Email: lfhgggg@163.com
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Abstract

Since embryonic stem cells (ESCs) were first identified, significant progress has been achieved. However, the establishment of buffalo ESCs (bESCs) is still unclear. This study was undertaken to explore the effect of the blastocyst stage on the isolation of bESCs. Firstly, our results indicated that the pluripotent genes were mainly expressed at the early stages of blastocyst, and the attachment and colony formation rates of bESCs derived from expanded blastocyst and hatched blastocyst were significantly higher than early blastocyst and blastocyst. In the meantime, bESCs showed positive alkaline phosphatase activity and expressed genes like OCT4, NANOG, SOX2, c-MYC, CDH1, KLF4, and TBX3. Immunofluorescence also confirmed the expression of OCT4, SOX2. Embryoid bodies expressing three marker genes were generated from the differentiation experiment, and fibroblast, epithelial, and neuron-like cells were induced. Moreover, naive-related genes KLF4, TBX3, primed-related genes FGF5, ACTA2 were expressed in the cells, but not REX-1. Immunofluorescence and western blot confirmed the FGF5 expression. Furthermore, bESCs could maintain pluripotency with the signal of LIF and bFGF. In summary, our results indicated that expanded blastocyst and hatched blastocyst are more suitable for bESCs isolation.

Keywords

buffalodifferent stages of blastocystembryonic stem cellsexpression pattern of pluripotent genes
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 28 , Issue 6 , December 2022 , pp. 2093 - 2104
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Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press on behalf of the Microscopy Society of America

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Footnotes

†

Xin Zhao and Qiuyan Ruan contributed equally to this work.

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