title: Does serum cause lipid-droplet accumulation in bovine embryos produced in vitro, during developmental days 1 to 4?
abstract: OBJECTIVE
  Serum supplementation has shown to have beneficial effects on in vitro bovine embryo development. However, it is often assumed that serum supplementation may produce mitochondrial damage and this damage would generate lipid accumulation, a major obstacle for cryopreservation. The aim of the present study is to investigate the previous assumptions in early embryonic stages.
  
  
  METHODS
  We considered in vitro produced bovine embryos from day 1 to 4 of development, which were grown in presence of serum from days 1, 2 or 3 or in absence of it. Electron transmission micrographs allowed us to quantify the area occupied by lipid droplets and by the different mitochondrial types to evaluate serum effect. Using confocal microscopy we analyzed mitochondrial activity and location.
  
  
  RESULTS
  We found no evidence of lipid droplets accumulation or mitochondrial degeneration or reduction of mitochondrial area in serum supplemented media. Further, our results suggest that events of mitochondrial proliferation are taking place even in serum supplemented media.
  
  
  CONCLUSIONS
  Serum does not produce lipid accumulation or mitochondrial damage in bovine embryos from 2 to 16 cells. When serum was added to embryo culture medium on day 3 of development, there were ultrastructural signs of a beneficial effect for embryo development. The lack of serum until day 3 may also avoid the unnecessary exposure to potentially inhibitory factors present on it.
authors.affiliation: Instituto Nacional de Parasitología "M. F. Chaben" ANLIS Malbrán, Av. Paseo Colón 568, Ciudad de Buenos Aires, Argentina, melcrocco@yahoo.com.
authors.forename: Melisa Candela
authors.initials: MC
authors.lastname: Crocco
authors.forename: Diana Mabel
authors.initials: DM
authors.lastname: Kelmansky
authors.forename: Marta Inés
authors.initials: MI
authors.lastname: Mariano
mesh_terms: D000818:Animals; D001947:Breeding / Q000379:methods; D002417:Cattle; D015925:Cryopreservation; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D046149:Embryo Culture Techniques / Q000379:methods*; D004622:Embryo, Mammalian / Q000187:drug effects; D047108:Embryonic Development; D005260:Female; D008055:Lipids; D008928:Mitochondria / Q000378:metabolism*; D011247:Pregnancy; D027724:Reproductive Techniques, Assisted / Q000662:veterinary*; D044967:Serum; D015053:Zygote
chemical_list: D016895:Culture Media, Serum-Free; D008055:Lipids
references.citation: Hum Reprod. 2000 Dec;15(12):2621-33
references.pmid: 11098036
references.citation: Biol Reprod. 2000 May;62(5):1459-65
references.pmid: 10775201
references.citation: New Phytol. 2003 Dec;160(3):463-478
references.pmid: 33873653
references.citation: Theriogenology. 1998 Jan 1;49(1):83-102
references.pmid: 10732123
references.citation: Mol Reprod Dev. 2005 Oct;72(2):201-7
references.pmid: 15968626
references.citation: J Cell Biol. 2003 Jan 20;160(2):189-200
references.pmid: 12527753
references.citation: Biol Reprod. 1995 Dec;53(6):1385-91
references.pmid: 8562695
references.citation: Proc Natl Acad Sci U S A. 1968 Oct;61(2):598-605
references.pmid: 4176482
references.citation: Reproduction. 2009 Apr;137(4):619-24
references.pmid: 19176592
references.citation: Biol Reprod. 1999 Jun;60(6):1345-52
references.pmid: 10330091
references.citation: Biol Reprod. 2010 Dec;83(6):909-18
references.pmid: 20686180
references.citation: Reproduction. 2012 Apr;143(4):417-27
references.pmid: 22408180
references.citation: Biol Reprod. 2000 Jun;62(6):1866-74
references.pmid: 10819794
references.citation: J Exp Bot. 2006;57(6):1225-43
references.pmid: 16556626
references.citation: Biol Reprod. 2001 May;64(5):1375-85
references.pmid: 11319141
references.citation: Hum Reprod Update. 1995 Mar;1(2):91-148
references.pmid: 15726768
references.citation: Stain Technol. 1964 Mar;39:111-4
references.pmid: 14127791
references.citation: Biol Reprod. 1999 Feb;60(2):534-9
references.pmid: 9916025
references.citation: Mol Reprod Dev. 2002 Feb;61(2):234-48
references.pmid: 11803560
references.citation: Theriogenology. 1995 Apr 15;43(6):1115-28
references.pmid: 16727698
references.citation: Zentralbl Veterinarmed A. 1994 May;41(4):307-16
references.pmid: 7975973
references.citation: Theriogenology. 1994;41(6):1241-9
references.pmid: 16727477
references.citation: Theriogenology. 1983 Oct;20(4):407-16
references.pmid: 16725857
references.citation: J Reprod Dev. 2011 Dec;57(6):681-9
references.pmid: 21804301
references.citation: Hum Reprod Update. 2010 Sep-Oct;16(5):488-509
references.pmid: 20231166
references.citation: J Assist Reprod Genet. 2013 Mar;30(3):293-303
references.pmid: 23430228
references.citation: Hum Reprod. 1996 Jan;11(1):177-83
references.pmid: 8671182
references.citation: Biol Reprod. 2010 Jul;83(1):52-62
references.pmid: 20130269
references.citation: Am J Physiol Endocrinol Metab. 2002 Aug;283(2):E226-32
references.pmid: 12110526
references.citation: J Cell Biol. 1970 Aug;46(2):405-8
references.pmid: 5449185
references.citation: Biol Reprod. 2010 Jan;82(1):76-85
references.pmid: 19696017
references.citation: Hum Reprod. 2010 Mar;25(3):768-78
references.pmid: 20007613
references.citation: J Reprod Fertil. 1999 Jul;116(2):373-8
references.pmid: 10615263
references.citation: Annu Rev Genet. 2005;39:503-36
references.pmid: 16285870
references.citation: Mol Hum Reprod. 2009 May;15(5):279-85
references.pmid: 19251762
references.citation: Mol Reprod Dev. 1999 Jul;53(3):325-35
references.pmid: 10369393
references.citation: Zygote. 2011 Nov;19(4):297-306
references.pmid: 21411040
references.citation: Biochim Biophys Acta. 2006 Feb;1762(2):140-7
references.pmid: 16054341
references.citation: Hum Reprod. 2000 Jul;15 Suppl 2:189-98
references.pmid: 11041524
references.citation: Acta Anat (Basel). 1986;125(1):27-31
references.pmid: 3953241
references.citation: Mol Reprod Dev. 2002 Jan;61(1):57-66
references.pmid: 11774376
references.citation: Theriogenology. 2003 Feb;59(3-4):889-99
references.pmid: 12517391
references.citation: J Reprod Dev. 2003 Jun;49(3):193-202
references.pmid: 14967928
references.citation: Theriogenology. 1998 Jan 1;49(1):103-14
references.pmid: 10732124
references.citation: Biol Reprod. 2011 Mar;84(3):572-80
references.pmid: 21076083
references.citation: Biol Reprod. 2003 Jan;68(1):236-43
references.pmid: 12493719
references.citation: Theriogenology. 2006 May;65(8):1551-62
references.pmid: 16229883
references.citation: Reproduction. 2004 Sep;128(3):269-80
references.pmid: 15333778
references.citation: Theriogenology. 2001 Mar 1;55(4):923-36
references.pmid: 11291915
references.citation: Dev Dyn. 1996 Jan;205(1):64-72
references.pmid: 8770552
references.citation: J Cell Biol. 1963 Apr;17:208-12
references.pmid: 13986422
references.citation: Hum Reprod Update. 2003 Nov-Dec;9(6):557-82
references.pmid: 14714592
references.citation: Mitochondrion. 2002 May;1(5):425-35
references.pmid: 16120295
references.citation: Biol Reprod. 1991 Nov;45(5):736-42
references.pmid: 1756211