Reprogramming of human somatic cells using human and animal oocytes

Young Chung, Colin E. Bishop, Nathan R. Treff, Stephen J. Walker, Vladislav M. Sandler, Sandy Becker, Irina Klimanskaya, Wan Song Wun, Randall Dunn, Rebecca M. Hall, Jing Su, Shi Jiang Lu, Marc Maserati, Young Ho Choi, Richard Scott, Anthony Atala, Ralph Dittman, Robert Lanza

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

There is renewed interest in using animal oocytes to reprogram human somatic cells. Here we compare the reprogramming of human somatic nuclei using oocytes obtained from animal and human sources. Comparative analysis of gene expression in morula-stage embryos was carried out using single-embryo transcriptome amplification and global gene expression analyses. Genomic DNA fingerprinting and PCR analysis confirmed that the nuclear genome of the cloned embryos originated from the donor somatic cell. Although the human-human, human-bovine, and human-rabbit clones appeared morphologically similar and continued development to the morula stage at approximately the same rate (39, 36, and 36%, respectively), the pattern of reprogramming of the donor genome was dramatically different. In contrast to the interspecies clones, gene expression profiles of the human-human embryos showed that there was extensive reprogramming of the donor nuclei through extensive upregulation, and that the expression pattern was similar in key upregulation in normal control embryos. To account for maternal gene expression, enucleated oocyte transcriptome profiles were subtracted from the corresponding morula-stage embryo profiles. t-Test comparisons (median-normalized data @ fc > 4; p < 0.005) between human in vitro fertilization (IVF) embryos and human-bovine or human-rabbit interspecies somatic cell transfer (iSCNT) embryos found between 2400 and 2950 genes that were differentially expressed, the majority (60-70%) of which were downregulated, whereas the same comparison between the bovine and rabbit oocyte profiles found no differences at all. In contrast to the iSCNT embryos, expression profiles of human-human clones compared to the age-matched IVF embryos showed that nearly all of the differentially expressed genes were upregulated in the clones. Importantly, the human oocytes significantly upregulated Oct-4, Sox-2, and nanog (22-fold, 6-fold, and 12-fold, respectively), whereas the bovine and rabbit oocytes either showed no difference or a downregulation of these critical pluripotency-associated genes, effectively silencing them. Without appropriate reprogramming, these data call into question the potential use of these discordant animal oocyte sources to generate patient-specific stem cells.

Original languageEnglish
Pages (from-to)213-223
Number of pages11
JournalCloning and Stem Cells
Volume11
Issue number2
DOIs
StatePublished - Jun 1 2009
Externally publishedYes

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Oocytes
Embryonic Structures
Morula
Clone Cells
Transcriptome
Rabbits
Fertilization in Vitro
Gene Expression
Up-Regulation
Down-Regulation
Tissue Donors
Genome
DNA Fingerprinting
Embryo Transfer
Gene Silencing
Genes
Stem Cells
Mothers

ASJC Scopus subject areas

  • Biotechnology
  • Developmental Biology

Cite this

Chung, Y., Bishop, C. E., Treff, N. R., Walker, S. J., Sandler, V. M., Becker, S., ... Lanza, R. (2009). Reprogramming of human somatic cells using human and animal oocytes. Cloning and Stem Cells, 11(2), 213-223. https://doi.org/10.1089/clo.2009.0004

Reprogramming of human somatic cells using human and animal oocytes. / Chung, Young; Bishop, Colin E.; Treff, Nathan R.; Walker, Stephen J.; Sandler, Vladislav M.; Becker, Sandy; Klimanskaya, Irina; Wun, Wan Song; Dunn, Randall; Hall, Rebecca M.; Su, Jing; Lu, Shi Jiang; Maserati, Marc; Choi, Young Ho; Scott, Richard; Atala, Anthony; Dittman, Ralph; Lanza, Robert.

In: Cloning and Stem Cells, Vol. 11, No. 2, 01.06.2009, p. 213-223.

Research output: Contribution to journalArticle

Chung, Y, Bishop, CE, Treff, NR, Walker, SJ, Sandler, VM, Becker, S, Klimanskaya, I, Wun, WS, Dunn, R, Hall, RM, Su, J, Lu, SJ, Maserati, M, Choi, YH, Scott, R, Atala, A, Dittman, R & Lanza, R 2009, 'Reprogramming of human somatic cells using human and animal oocytes', Cloning and Stem Cells, vol. 11, no. 2, pp. 213-223. https://doi.org/10.1089/clo.2009.0004
Chung Y, Bishop CE, Treff NR, Walker SJ, Sandler VM, Becker S et al. Reprogramming of human somatic cells using human and animal oocytes. Cloning and Stem Cells. 2009 Jun 1;11(2):213-223. https://doi.org/10.1089/clo.2009.0004
Chung, Young ; Bishop, Colin E. ; Treff, Nathan R. ; Walker, Stephen J. ; Sandler, Vladislav M. ; Becker, Sandy ; Klimanskaya, Irina ; Wun, Wan Song ; Dunn, Randall ; Hall, Rebecca M. ; Su, Jing ; Lu, Shi Jiang ; Maserati, Marc ; Choi, Young Ho ; Scott, Richard ; Atala, Anthony ; Dittman, Ralph ; Lanza, Robert. / Reprogramming of human somatic cells using human and animal oocytes. In: Cloning and Stem Cells. 2009 ; Vol. 11, No. 2. pp. 213-223.
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