Please use this identifier to cite or link to this item: https://doi.org/10.1371/journal.pone.0097974
Title: Karyotype characterization of in vivo- And in vitro-derived porcine parthenogenetic cell lines
Authors: Liu Q.
Zhang M.
Hou D.
Han X.
Jin Y.
Zhao L.
Nie X. 
Zhou X.
Yun T.
Zhao Y.
Huang X.
Hou D.
Yang N.
Wu Z.
Li X.
Li R.
Keywords: animal cell
article
blastocyst
blastoma
cell line
chromosome G band
controlled study
down regulation
embryo
embryo transfer
embryonic stem cell
fetus
fibroblast
flow cytometry
genome imprinting
haploidy
in vitro study
in vivo study
karyotype
microarray analysis
mouse
nonhuman
polyploidy
swine
animal
cell culture
cell line
chromosome banding pattern
cluster analysis
gene expression profiling
gene expression regulation
genetics
haploidy
karyotyping
metabolism
oocyte
parthenogenesis
Animals
Blastocyst
Blastomeres
Cell Line
Cells, Cultured
Chromosome Banding
Cluster Analysis
Embryo Transfer
Embryonic Stem Cells
Fetus
Fibroblasts
Flow Cytometry
Gene Expression Profiling
Gene Expression Regulation, Developmental
Haploidy
In Vitro Techniques
Karyotype
Karyotyping
Oocytes
Parthenogenesis
Swine
Issue Date: 2014
Citation: Liu Q., Zhang M., Hou D., Han X., Jin Y., Zhao L., Nie X., Zhou X., Yun T., Zhao Y., Huang X., Hou D., Yang N., Wu Z., Li X., Li R. (2014). Karyotype characterization of in vivo- And in vitro-derived porcine parthenogenetic cell lines. PLoS ONE 9 (5) : e97974. ScholarBank@NUS Repository. https://doi.org/10.1371/journal.pone.0097974
Rights: Attribution 4.0 International
Abstract: Mammalian haploid cell lines provide useful tools for both genetic studies and transgenic animal production. To derive porcine haploid cells, three sets of experiments were conducted. First, genomes of blastomeres from 8-cell to 16-cell porcine parthenogenetically activated (PA) embryos were examined by chromosome spread analysis. An intact haploid genome was maintained by 48.15% of blastomeres. Based on this result, two major approaches for amplifying the haploid cell population were tested. First, embryonic stem-like (ES-like) cells were cultured from PA blastocyst stage embryos, and second, fetal fibroblasts from implanted day 30 PA fetuses were cultured. A total of six ES-like cell lines were derived from PA blastocysts. No chromosome spread with exactly 19 chromosomes (the normal haploid complement) was found. Four cell lines showed a tendency to develop to polyploidy (more than 38 chromosomes). The karyotypes of the fetal fibroblasts showed different abnormalities. Cells with 19-38 chromosomes were the predominant karyotype (59.48-60.91%). The diploid cells were the second most observed karyotype (16.17%-22.73%). Although a low percentage (3.45-8.33%) of cells with 19 chromosomes were detected in 18.52% of the fetus-derived cell lines, these cells were not authentic haploid cells since they exhibited random losses or gains of some chromosomes. The haploid fibroblasts were not efficiently enriched via flow cytometry sorting. On the contrary, the diploid cells were efficiently enriched. The enriched parthenogenetic diploid cells showed normal karyotypes and expressed paternally imprinted genes at extremely low levels. We concluded that only a limited number of authentic haploid cells could be obtained from porcine cleavage-stage parthenogenetic embryos. Unlike mouse, the karyotype of porcine PA embryo-derived haploid cells is not stable, long-term culture of parthenogenetic embryos, either in vivo or in vitro, resulted in abnormal karyotypes. The porcine PA embryo-derived diploid fibroblasts enriched from sorting might be candidate cells for paternally imprinted gene research. © 2014 Liu et al.
Source Title: PLoS ONE
URI: https://scholarbank.nus.edu.sg/handle/10635/161409
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0097974
Rights: Attribution 4.0 International
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