Inefficient reprogramming of the hematopoietic stem cell genome following nuclear transfer.
In general, cloning undifferentiated preimplantation embryos (blastomeres) or embryonic stem cells is more efficient than cloning differentiated somatic cells. Therefore, there has been an assumption that tissue-specific stem cells might serve as efficient donors for nuclear transfer because of the undifferentiated state of their genome. Here, we show that this is not the case with adult hematopoietic stem cells (HSCs). Although we have demonstrated for the first time that mouse HSCs can be cloned to generate offspring, the birth rates (0-0.7%) were lowest among the clones tested (cumulus, immature Sertoli and fibroblast cells). Only 6% of reconstructed embryos reached the morula or blastocyst stage in vitro (versus 46% for cumulus clones; P < 5 x 10(-10)). Transcription and gene expression analyses of HSC clone embryos revealed that they initiated zygotic gene activation (ZGA) at the appropriate timing, but failed to activate five out of six important embryonic genes examined, including Hdac1 (encoding histone deacetylase 1), a key regulator of subsequent ZGA. These results suggest that the HSC genome has less plasticity than we imagined, at least in terms of reprogrammability in the ooplasm after nuclear transfer.
Inoue
Kimiko
K
RIKEN Bioresource Center, Tsukuba, Ibaraki 305-0074, Japan.
Ogonuki
Narumi
N
Miki
Hiromi
H
Hirose
Michiko
M
Noda
Shinichi
S
Kim
Jin-Moon
JM
Aoki
Fugaku
F
Miyoshi
Hiroyuki
H
Ogura
Atsuo
A
D000818:Animals; D001755:Blastocyst / Q000166:cytology; D002467:Cell Nucleus / Q000235:genetics; D019976:Cloning, Organism / Q000379:methods*; D004624:Embryo Transfer; D005260:Female; D015870:Gene Expression; D016678:Genome; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D056284:Histone Deacetylase 1; D006655:Histone Deacetylases / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008811:Mice, Inbred DBA; D053652:Nuclear Transfer Techniques; D015533:Transcriptional Activation / Q000235:genetics
C484389:Hdac1 protein, mouse; D056284:Histone Deacetylase 1; D006655:Histone Deacetylases