Centrosome amplification and chromosomal instability in human and animal parthenogenetic cell lines.
Parthenotes have been proposed as a source of embryonic stem cells but they lack the centriole which is inherited through the sperm in all mammalian species, except for rodents. We investigated the centrosome of parthenotes and parthenogenetic embryonic stem cells using parthenogenetic and biparental pig pre-implantation embryos, human and pig parthenogenetic and biparental embryonic stem cells, sheep fibroblasts derived from post implantation parthenogenetic and biparental embryos developed in vivo. We also determined the level of aneuploidy in parthenogenetic cells. Oocytes of all species were activated using ionomycin and 6-dimethylaminopurine (6-DMAP). Over 60% of parthenogenetic blastomeres were affected by an excessive number of centrioles. Centrosome amplification, was observed by microscopical and ultrastructural analysis also in parthenogenetic cell lines of all three species. Over expression of PLK2 and down regulation of CCNF, respectively involved in the stimulation and inhibition of centrosome duplication, were present in all species. We also detected down regulation of spindle assembly checkpoint components such as BUB1, CENPE and MAD2. Centrosome amplification was accompanied by multipolar mitotic spindles and all cell lines were affected by a high rate of aneuploidy. These observations indicate a link between centrosome amplification and the high incidence of aneuploidy and suggest that parthenogenetic stem cells may be a useful model to investigate how aneuploidy can be compatible with cell proliferation and differentiation.
Laboratory of Biomedical Embryology, Centre for Stem Cell Research (UniStem), Università degli Studi di Milano, Via Celoria 10, 20133 Milan, Italy. tiziana.brevini@unimi.it
Tiziana A L
TA
Brevini
Georgia
G
Pennarossa
Sara
S
Maffei
Gianluca
G
Tettamanti
Arianna
A
Vanelli
Sara
S
Isaac
Amir
A
Eden
Sergio
S
Ledda
Magda
M
de Eguileor
Fulvio
F
Gandolfi
D000782:Aneuploidy; D000818:Animals; D001757:Blastomeres / Q000378:metabolism* / Q000473:pathology; D061207:Calcium Ionophores / Q000009:adverse effects / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D002460:Cell Line; D018385:Centrosome / Q000378:metabolism* / Q000473:pathology; D043171:Chromosomal Instability; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D006801:Humans; D015759:Ionomycin / Q000009:adverse effects / Q000494:pharmacology; D064246:Mad2 Proteins; D009865:Oocytes / Q000378:metabolism / Q000473:pathology; D010312:Parthenogenesis; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D012756:Sheep; D008941:Spindle Apparatus / Q000378:metabolism* / Q000473:pathology; D013552:Swine
D061207:Calcium Ionophores; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; C577697:MAD2L1 protein, human; D064246:Mad2 Proteins; D012097:Repressor Proteins; C068688:centromere protein E; D015759:Ionomycin; C584277:BUB1 protein, human; C090464:Bub1 spindle checkpoint protein; D017346:Protein Serine-Threonine Kinases