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Using Mouse Oocytes to Assess Human Gene Function During Meiosis I
Published on: April 10, 2018
Reprogramming of human somatic cells using human and animal oocytes
Young Chung1, Colin E Bishop, Nathan R Treff
1Advanced Cell Technology, Worcester, MA 01605, USA. rlanza@advancedcell.com
Cloning and Stem Cells
|February 4, 2009
Summary
Animal oocytes show poor reprogramming of human somatic cells compared to human oocytes. This difference in gene expression, particularly for pluripotency genes, questions the use of animal oocytes for patient-specific stem cell generation.
Area of Science:
- Reproductive Biology
- Stem Cell Biology
- Genomics
Background:
- Renewed interest exists in using animal oocytes for human somatic cell reprogramming.
- Successful reprogramming is crucial for generating patient-specific stem cells.
Purpose of the Study:
- To compare the reprogramming efficiency of human somatic nuclei using human versus animal oocytes.
- To analyze gene expression patterns in embryos derived from interspecies and intraspecies somatic cell transfer.
Main Methods:
- Somatic cell nuclear transfer into human, bovine, and rabbit oocytes.
- Single-embryo transcriptome amplification and global gene expression analysis.
- Genomic DNA fingerprinting and PCR analysis for nuclear origin confirmation.
- Differential gene expression analysis using t-tests, accounting for maternal gene expression.
Main Results:
- Human-human, human-bovine, and human-rabbit clones developed to the morula stage at similar rates.
- Gene expression reprogramming was significantly different: human oocytes extensively upregulated pluripotency genes (Oct-4, Sox-2, Nanog), while animal oocytes showed no difference or downregulation.
- Interspecies clones (human-bovine, human-rabbit) exhibited significant gene downregulation compared to human-human clones and in vitro fertilization (IVF) embryos.
Conclusions:
- Animal oocytes, particularly bovine and rabbit, are inefficient at reprogramming human somatic cell nuclei.
- The failure to upregulate key pluripotency genes in interspecies clones suggests they are not effectively reprogrammed.
- These findings question the utility of animal oocytes for generating patient-specific induced pluripotent stem cells due to discordant reprogramming.
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