Page 1 of 41Reproduction Advance Publication first posted on 27 March 2015 as Manuscript REP-15-0048
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Title: Cheetah interspecific SCNT followed by embryo aggregation improves in vitro
2
development but not pluripotent gene expression
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Short title: SCNT and embryo aggregation in felids
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Authors and affiliations: Moro L.N.1,2, Hiriart M.I.1,2, Buemo C.1,2, Jarazo J.1, Sestelo A.3,
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Veraguas D.4 Rodriguez-Alvarez L.4 and Salamone D.F.1,2*
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*Correspondence: Daniel F. Salamone, Laboratory of Animal Biotechnology, Agriculture
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Faculty, University of Buenos Aires, Av. San Martin 4453 (1417), Buenos Aires,
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Argentina. E-mail: [email protected].
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1
Laboratory of Animal Biotechnology, Agriculture Faculty, University of Buenos Aires,
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Av. San Martin 4453 (1417), Buenos Aires, Argentina.
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2
12
(C1033AAJ), Buenos Aires, Argentina.
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3
14
República de la India 3000, Buenos Aires, Argentina.
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4
16
Vicente Méndez 595, Chillán, Chile.
National Institute of Scientific and Technological Research, Av. Rivadavia 1917
Laboratory of Reproductive Biotechnology, Zoological Garden of Buenos Aires,
Department of Animal Science, Veterinary Faculty, University of Concepción, Av.
17 18 19 20 21 22 23 1
Copyright © 2015 by the Society for Reproduction and Fertility.
Page 2 of 41
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ABSTRACT
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The aim of this study was to evaluate the capacity of domestic cat (Dc, Felis silvestris)
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oocytes to reprogram the nucleus of cheetah (Ch, Acinonyx jubatus) cells by inter-species
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SCNT (iSCNT), by using embryo aggregation. Domestic cat oocytes were in vitro matured
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and subjected to zona pellucida free (ZP-free) SCNT or iSCNT, depending on whether the
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nucleus donor cell was of domestic cat or cheetah, respectively. ZP-free reconstructed
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embryos were then cultured in microwells individually (Dc1X and Ch1X groups) or in
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couples (Dc2X and Ch2X groups). Embryo aggregation improved in vitro development
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obtaining 27.4%, 47.7%, 16.7% and 28.3% of blastocyst rates in the Dc1X, Dc2X, Ch1X
34
and Ch2X groups, respectively (p
1
Title: Cheetah interspecific SCNT followed by embryo aggregation improves in vitro
2
development but not pluripotent gene expression
3
Short title: SCNT and embryo aggregation in felids
4
Authors and affiliations: Moro L.N.1,2, Hiriart M.I.1,2, Buemo C.1,2, Jarazo J.1, Sestelo A.3,
5
Veraguas D.4 Rodriguez-Alvarez L.4 and Salamone D.F.1,2*
6
*Correspondence: Daniel F. Salamone, Laboratory of Animal Biotechnology, Agriculture
7
Faculty, University of Buenos Aires, Av. San Martin 4453 (1417), Buenos Aires,
8
Argentina. E-mail: [email protected].
9
1
Laboratory of Animal Biotechnology, Agriculture Faculty, University of Buenos Aires,
10
Av. San Martin 4453 (1417), Buenos Aires, Argentina.
11
2
12
(C1033AAJ), Buenos Aires, Argentina.
13
3
14
República de la India 3000, Buenos Aires, Argentina.
15
4
16
Vicente Méndez 595, Chillán, Chile.
National Institute of Scientific and Technological Research, Av. Rivadavia 1917
Laboratory of Reproductive Biotechnology, Zoological Garden of Buenos Aires,
Department of Animal Science, Veterinary Faculty, University of Concepción, Av.
17 18 19 20 21 22 23 1
Copyright © 2015 by the Society for Reproduction and Fertility.
Page 2 of 41
24
ABSTRACT
25
26
The aim of this study was to evaluate the capacity of domestic cat (Dc, Felis silvestris)
27
oocytes to reprogram the nucleus of cheetah (Ch, Acinonyx jubatus) cells by inter-species
28
SCNT (iSCNT), by using embryo aggregation. Domestic cat oocytes were in vitro matured
29
and subjected to zona pellucida free (ZP-free) SCNT or iSCNT, depending on whether the
30
nucleus donor cell was of domestic cat or cheetah, respectively. ZP-free reconstructed
31
embryos were then cultured in microwells individually (Dc1X and Ch1X groups) or in
32
couples (Dc2X and Ch2X groups). Embryo aggregation improved in vitro development
33
obtaining 27.4%, 47.7%, 16.7% and 28.3% of blastocyst rates in the Dc1X, Dc2X, Ch1X
34
and Ch2X groups, respectively (p
