Recombinant human albumin supports development of somatic cell nuclear transfer embryos in mice: toward the establishment of a chemically defined cloning protocol.
Culturing embryos in different media is a useful approach to characterize their nature in regard to "memory" of the donor nucleus and its "reprogramming" after somatic cell nuclear transfer (SCNT). However, efforts to elucidate the mechanisms of reprogramming are seriously undermined when embryo culture conditions are not completely defined. Using recombinant human albumin (rHA) is a step toward establishing defined culture conditions for mouse cloning. Recombinant HA supports blastocyst formation of cumulus cell-derived clones at a rate comparable with two types of bovine serum albumin (BSA); following transfer of blastocysts to the genital tract, rates of development to midgestation (10.5 dpc) were indistinguishable. rHA also supports the derivation of germline competent embryonic stem (ES) cells from SCNT blastocysts at a substantial rate compared with BSA counterparts and with zygotic blastocysts. Unlike the developmental parameters, the gene expression patterns of clones cultured in rHA or BSA were not superimposed; identical patterns were observed for zygotic blastocysts in the two albumins. In summary, the present study demonstrates that (1) rHA can replace BSA, proving a defined protein source for SCNT in mice; (2) although using rHA is similar to BSA, it is not equal (rHA leaves a mark on gene expression of clones but not zygotes). Future studies that investigate reprogramming after SCNT will need to consider not only the implications of culture media for cloning but also the supplement choice.
Max Planck Institute for Molecular Biomedicine, Münster, Germany.
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F
Cavaleri
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Gentile
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HR
Schöler
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M
Boiani
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology; D002478:Cells, Cultured; D019976:Cloning, Organism / Q000379:methods*; D003470:Culture Media; D046149:Embryo Culture Techniques; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000187:drug effects*; D047108:Embryonic Development; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D053652:Nuclear Transfer Techniques; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012709:Serum Albumin / Q000494:pharmacology*; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology; D014158:Transcription, Genetic
D003470:Culture Media; D011994:Recombinant Proteins; D012709:Serum Albumin; D012710:Serum Albumin, Bovine