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J Reprod Dev. 2017 Dec 15;63(6):611-616. doi: 10.1262/jrd.2017-074. Epub 2017 Aug 19.

Haploid embryonic stem cell lines derived from androgenetic and parthenogenetic rat blastocysts.

Author information

1
Center for Genetic Analysis of Behavior, National Institute for Physiological Sciences, Aichi 444-8787, Japan.
2
School of Life Science, The Graduate University for Advanced Studies, Aichi 444-8787, Japan.
3
Department of Physiology, Medical College of Wisconsin, WI 53226, USA.
4
Present: Center for Molecular Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
5
Graduate School of Science and Technology, Shinshu University, Nagano 386-8567, Japan.
6
Faculty of Textile Science and Technology, Shinshu University, Nagano 386-8567, Japan.
7
Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan.
8
Stanford University School of Medicine, CA 94305, USA.

Abstract

The present study was conducted to establish haploid embryonic stem (ES) cell lines using fluorescent marker-carrying rats. In the first series, 7 ES cell lines were established from 26 androgenetic haploid blastocysts. However, only 1 ES cell line (ahES-2) was found to contain haploid cells (1n = 20 + X) by fluorescence-activated cell sorting (FACS) and karyotypic analyses. No chimeras were detected among the 10 fetuses and 41 offspring derived from blastocyst injection with the FACS-purified haploid cells. In the second series, 2 ES cell lines containing haploid cells (13% in phES-1 and 1% in phES-2) were established from 2 parthenogenetic haploid blastocysts. Only the phES-2 cell population was purified by repeated FACS to obtain 33% haploid cells. Following blastocyst injection with the FACS-purified haploid cells, no chimera was observed among the 11 fetuses; however, 1 chimeric male was found among the 47 offspring. Although haploid rat ES cell lines can be established from both blastocyst sources, FACS purification may be necessary for maintenance and chimera production.

KEYWORDS:

Blastocyst injection; FACS purification; Haploid ES cells; Rat blastocysts

PMID:
28824040
PMCID:
PMC5735273
DOI:
10.1262/jrd.2017-074
[Indexed for MEDLINE]
Free PMC Article

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