Variable reprogramming of the pluripotent stem cell marker Oct4 in mouse clones: distinct developmental potentials in different culture environments.
A prevailing view of cloning by somatic-cell nuclear transfer is that reprogramming of gene expression occurs during the first few hours after injection of the nucleus into an oocyte, that the process is stochastic, and that the type of reprogramming needed for cloning success is foreign and unlikely to be readily achieved in the ooplasm. Here, we present evidence that the release of reprogramming capacity is contingent on the culture environment of the clone while the contribution of aneuploidy to altered gene expression is marginal. In particular, the rate of blastocyst formation in clones and the regional distribution of mRNA for the pluripotent stem cell marker Oct4 in clonal blastocysts was highly dependent on the culture environment after cumulus cell nuclear transfer, unlike that in genetically equivalent zygotes. Epigenetic modifications of genetically identical somatic nuclei continue after the first cell division of the clones and are amenable to a degree of experimental control, and their development to the blastocyst stage and appropriate expression of Oct4 predict further outcome, such as derivation of embryonic stem (ES) cells, but not fetal development. This observation indicates that development to the blastocyst stage is not equivalent to full reprogramming and lends support to the novel concept that ES cells are not the equivalent of the inner cell mass, hence the discrepancy between ES cell derivability and fetal development of clones.
Boiani
Michele
M
Max Planck Institute for Molecular Biomedicine, Mendelstrasse 7, D-48149 Münster, Germany. mboiani@mpi-muenster.mpg.de
Gentile
Luca
L
Gambles
Vivian V
VV
Cavaleri
Fatima
F
Redi
Carlo A
CA
Schöler
Hans R
HR
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D002467:Cell Nucleus / Q000235:genetics; D019976:Cloning, Organism; D003470:Culture Media / Q000737:chemistry*; D046149:Embryo Culture Techniques; D005260:Female; D005786:Gene Expression Regulation; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D053652:Nuclear Transfer Techniques; D050814:Octamer Transcription Factor-3 / Q000235:genetics* / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D011786:Quality Control; D012333:RNA, Messenger / Q000378:metabolism; D015053:Zygote / Q000166:cytology / Q000378:metabolism
D003470:Culture Media; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D012333:RNA, Messenger