Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Journal of Assisted Reproduction and Genetics
Published in: Journal of Assisted Reproduction and Genetics 10/2013

01-10-2013 | Embryo Biology

Does serum cause lipid-droplet accumulation in bovine embryos produced in vitro, during developmental days 1 to 4?

Authors: Melisa Candela Crocco, Diana Mabel Kelmansky, Marta Inés Mariano

Published in: Journal of Assisted Reproduction and Genetics | Issue 10/2013

Login to get access

Summary

Purpose

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.
Literature
1.
Abe H, Hoshi H. Evaluation of bovine embryos produced in high performance serum-free media. J Reprod Dev. 2003;49(3):193–202.PubMedCrossRef
2.
Abe H, Yamahita S, Itoh T, Satoh T, Hoshi H. Ultrastructure of bovine embryos developed from in vitro-matured and –fertilized oocytes: Comparative morphological evaluation of embryos cultured either in serum-free medium or in serum-supplemented medium. Mol Reprod Dev. 1999;53:325–35.PubMedCrossRef
3.
Abe H, Yamashita S, Satoh T, Hoshi H. Accumulation of cytoplasmic lipid droplets in bovine embryos and cryotolerance of embryos developed in different culture systems using serum-free or serum-containing media. Mol Reprod Dev. 2002;61:57–66.PubMedCrossRef
4.
Algriany O, Vos PLAM, Sirard MA, Dieleman SJ. Switch in the expression of genes involved in lipid metabolism for in vivo matured bovine oocytes and blastocysts. Reprod Fert Devel. 2007;19:244–4.
5.
Barnett DK, Bavister BD. Inhibitory effect of glucose and phosphate on the second cleavage division of hamster embryos: is it linked to metabolism? Hum Reprod. 1996;11(l):177–83.PubMedCrossRef
6.
Barnett DK, Kimura J, Bavister BD. Translocation of active mitochondria during hamster preimplantation embryo development studied by confocal laser scanning microscopy. Developmental Dynamics. 1996;205:64–72.PubMedCrossRef
7.
Bavister BD. Culture of preimplantation embryos: facts and artifacts. Hum Reprod Update. 1995;1:91–148.PubMedCrossRef
8.
Bavister BD, Squirrell JM. Mitochondrial distribution and function in oocytes and early embryos. Hum Reprod. 2000;15 Suppl 2:189–98.PubMedCrossRef
9.
Camargo O, Ruiz T, Olivera M. Theoretical model for explaining accumulation of fat drops in in vitro produced bovine embryos. Acta Biológica Colombiana. 2008;13(2):89–102.
10.
Carolan C, Lonergan P, Van Langendonckt A, Mermillod P. Factors affecting bovine embryo development in synthetic oviduct fluid following oocyte maturation and fertilization in vitro. Theriogenology. 1995;43:1115–28.PubMedCrossRef
11.
Chen H, Detmer SA, Ewald AJ, Griffin EE, Fraser SE, Chan DC. Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development. The Journal of Cell Biology. 2003;160(2):189–200.PubMedCrossRefPubMedCentral
12.
Chi MM-Y, Hoehn A, Moley KH. Metabolic changes in the glucose-induced apoptotic blastocyst suggest alterations in mitochondrial physiology. Am J Physiol Endocrinol Metab. 2002;283:E226–32.PubMed
13.
Chiaratti MR, Bressan FF, Ferreira CR, Caetano AR, Smith LC, Vercesi AE, et al. Embryo mitochondrial DNA depletion is reversed during early embryogenesis in cattle. Biol Reprod. 2010;82:76–85.PubMedCrossRef
14.
Crocco M, Lauría L, Alberio R, Mariano MI (2011) Effect of serum on the mitochondrial active area on developmental days 1 to 4 in in vitro-produced bovine embryos. Zygote 19(4):297–306.
15.
Crosier AE, Farin PW, Dykstra MJ, Alexander JE, Farin CE. Ultrastructural morphometry of bovine compact murulae produced in vivo or in vitro. Biol Reprod. 2000;62:1459–65.PubMedCrossRef
16.
Crosier AE, Farin PW, Dykstra MJ, Alexander JE, Farin CE. Ultrastructural morphometry of bovine blastocysts produced in vivo or in vitro. Biol Reprod. 2001;64:1375–85.PubMedCrossRef
17.
Diez C, Heyman Y, Le Bourhis D, Guyader-Joly C, Degrouard J, Renard JP. Delipidating in vitro-produced bovine zygotes: Effect on further development and consequences for freezability. Theriogenology. 2001;55:923–36.PubMedCrossRef
18.
Dorland M, Gardner DK, Trounson AO. Serum in synthetic oviduct fluid causes mitochondrial degeneration in ovine embryos. J Reprod Fertil Abstract Series. 1994;13:70.
19.
Dunning KR, Cashman K, Russell DL, Thompson JG, Norman RJ, Robker RL. Beta-oxidation is essential for mouse oocyte developmental competence and early embryo development. Biol Reprod. 2010;83(6):909–18.PubMedCrossRef
20.
Duque P, Gómez E, Díaz E, Facal N, Hidalgo C, Díez C. Use of two replacements of serum during bovine embryo culture in vitro. Theriogenology. 2003;59(3–4):889–99.PubMedCrossRef
21.
Ferguson EM, Leese HJ. Triglyceride content of bovine oocytes and early embryos. J Reprod Fertil. 1999;116(2):373–8.PubMedCrossRef
22.
Gardner DK. Changes in requirements and utilization of nutrients during mammalian preimplantation embryo development and their significance in embryo culture. Theriogenology. 1998;49:83–102.PubMedCrossRef
23.
Gardner DK, Lane M, Stevens J, Schoolcraft WB. Ongoing development of a human blastocyst cultura system. Fertility and Sterility. 2002;78:S8.CrossRef
24.
Guyader-Joly C. Activité métabolique et aptitude à la congélation de l’embryon bovin produit in vitro. Elevage et Insémination. 1998;288:3–23.
25.
Hackenbrock CR. Chemical and physical fixation of isolated mitochondria in low-energy and high-energy states. Proceedings of the National Academy of Sciences USA. 1968;61:598–605.CrossRef
26.
Hyttel P, Lehn-Jensen H, Greve T. Ultrastructure of bovine embryos frozen and thawed by a two-step freezing method. Acta anat. 1986;125:27–31.PubMedCrossRef
27.
Kimura K, Spate LD, Green MP, Roberts RM. Effects of D-glucose concentration, D-fructose, and inhibitors of enzymes of the pentose phosphate pathway on the development and sex bovine blastocysts. Mol Reprod Dev. 2005;72:201–7.PubMedCrossRef
28.
Krisher RL, Bavister BD. Responses of oocytes and embryos to the culture environment. Theriogenology. 1998;49:103–14.PubMedCrossRef
29.
Krisher RL, Lane M, Bavister BD. Influence of semi-defined and defined culture media on the development, cell number and metabolism of bovine embryos. Theriogenology. 1998;49:207.CrossRef
30.
Krisher RL, Lane M, Bavister BD. Developmental competence and metabolism of bovine embryos cultured in semi-defined and defined culture media. Biol Reprod. 1999;60:1345–52.PubMedCrossRef
31.
32.
Leroy JLMR, Van Hoeck V, Clemente M, Rizos D, Gutierrez-Adan A, Van Soom A, et al. The effect of nutritionally induced hyperlipidaemia on in vitro bovine embryo quality. Hum Reprod. 2010;25(3):768–78.PubMedCrossRef
33.
34.
35.
36.
Mannella CA. The relevance of mitochondrial membrane topology to mitochondrial function. Biochimica et Biophysica Acta. 2006;1762:140–7.PubMedCrossRef
37.
Margineantu DH, Cox WG, Sundell L, Sherwood SW, Beechemb JM, Capaldi RA. Cell cycle dependent morphology changes and associated mtDNA redistribution in mitochondria of human cell lines. Mitochondrion. 2002;1:425–35.PubMedCrossRef
38.
Maurer HR. Towards serum-free, chemical defined media for mammalian cell culture. In: Freshney RI, editor. Animal Cell Culture: A Practical Approach. 2nd ed. Oxford: Oxford Univ Press; 1992. p. 15–46.
39.
40.
Mollenhauer HH. Plastic embedding mixtures for use in electron microscopy. Stain Technology. 1964;39:111.PubMed
41.
Momozawa K, Fukuda Y. Establishment of an advanced chemically defined medium for early embryos derived from in vitro matured and fertilized bovine oocytes. J Reprod Dev. 2011;57(6):681–9.PubMedCrossRef
42.
Mucci N, Aller J, Kaiser GG, Hozbor F, Cabodevila J, Alberio RH. Effect of estrous cow serum during bovine embryo culture on blastocyst development and cryotolerance after slow freezing or vitrification. Theriogenology. 2006;65(8):1551–62.PubMedCrossRef
43.
Okamoto K, Shaw JM. Mitochondrial morphology and dynamics in yeast and multicellular eukaryotes. Annual Review of Genetics. 2005;39:503–36.PubMedCrossRef
44.
Pinyopummintr T, Bavister BD. In vitro-matured/in vitro-fertilized bovine oocytes can develop into morulae/blastocysts in chemically defined protein-free cultured media. Biol Reprod. 1991;45:736–42.PubMedCrossRef
45.
Pinyopummintr T, Bavister BD. Development of bovine embryos in a cell-free culture medium: effects of type of serum, timing of its inclusion and heat inactivation. Theriogenology. 1994;41:1241–9.PubMedCrossRef
46.
47.
Rexroad Jr CE. Co-culture of domestic animal embryos. Theriogenology. 1989;31:105–14.CrossRef
48.
49.
Rieger D. Relationship between energy metabolism and development of early mammalian embryos. Theriogenology. 1992;37(1):75–93.CrossRef
50.
Rizos D, Gutiérrez-Adán A, Pérez-Garnelo S, De La Fuente J, Boland MP, Lonergan P. Bovine embryo culture in the presence or absence of serum: implications for blastocyst development, cryotolerance, and messenger RNA expression. Biol Reprod. 2003;68:236–43.PubMedCrossRef
51.
Rizos D, Ward F, Duffy P, Boland MP, Lonergan P. Consequences of bovine oocyte maturation, fertilization or early embryo development in vitro versus in vivo: Implications for blastocyst yield and blastocyst quality. Mol Reprod Dev. 2002;61:234–48.PubMedCrossRef
52.
Sata R, Tsuji H, Abe H, Yamashita S, Hoshi H. Fatty acid composition of bovine embryos cultured in serum-free and serum-containing medium during early embryonic development. J Reprod Dev. 1999;45:97–103.CrossRef
53.
54.
Shamsuddin M, Rodriguez-Martinez H. Fine structure of bovine blastocysts developed either in serum-free medium or in conventional co-culture with oviduct epithelial cells. J Vet Med. 1994;41:307–16.CrossRef
55.
St. John JC, Facucho-Oliveira J, Jiang Y, Kelly R, Salah R. Mitochondrial DNA transmission, replication and inheritance: a journey from the gamete through the embryo and into offspring and embryonic stem cells. Hum Reprod Update. 2010;16(5):488–509.PubMedCrossRef
56.
Summers MC, Biggers JD. Chemically defined media and the culture of mammalian preimplantation embryos: historical perspective and current issues. Hum Reprod Update. 2003;9(6):557–82.PubMedCrossRef
57.
Thompson JG, Gardner DK, Pugh PA, McMillan WH, Tervit HR. Lamb birth weight is affected by culture system utilized during in vitro pre-elongation development of ovine embryos. Biol Reprod. 1995;53:1385–91.PubMedCrossRef
58.
Trimarchi JR, Liu L, Portereld DM, Smith PJ, Keefe DL. Oxidative phosphorylation-dependent and -independent oxygen consumption by individual preimplantation mouse embryos. Biol Reprod. 2000;62:1866–74.PubMedCrossRef
59.
Ushijima H, Yamakawa H, Nagashima H. Cryopreservation of bovine pre-morula-stage in vitro matured/in vitro fertilized embryos after delipidation and before use in nucleus transfer. Biol Reprod. 1999;60:535–9.CrossRef
60.
Van Blerkom J. Mitochondria in human oogenesis and preimplantation embryogenesis: engines of metabolism, ionic regulation and developmental competence. Reproduction. 2004;128:269–80.PubMedCrossRef
61.
Van Blerkom J, Davis P, Alexander S. Differential mitochondrial inheritance between blastomeres in cleavage stage human embryos: determination at the pronuclear stage and relationship to microtubular organization, ATP content and developmental competence. Hum Reprod. 2000;15:2621–33.PubMedCrossRef
62.
63.
Wakefield SL, Lane M, Mitchell M. Impaired mitochondrial function in the preimplantation embryo perturbs fetal and placental development in the mouse. Biol Reprod. 2011;84(3):572–80.PubMedCrossRef
64.
Wilding M, Coppola G, Dale B, Di Matteo L. Mitochondria and human preimplantation embryo development. Reproduction. 2009;137:619–24.PubMedCrossRef
65.
Zhou G-B, Li N. Cryopreservation of porcine oocytes: recent advances. Mol Hum Reprod. 2009;15(5):279–85.PubMedCrossRef
Metadata
Title
Does serum cause lipid-droplet accumulation in bovine embryos produced in vitro, during developmental days 1 to 4?
Authors
Melisa Candela Crocco
Diana Mabel Kelmansky
Marta Inés Mariano
Publication date
01-10-2013
Publisher
Springer US
Published in
Journal of Assisted Reproduction and Genetics / Issue 10/2013
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI
https://doi.org/10.1007/s10815-013-0060-8

Other articles of this Issue 10/2013

Journal of Assisted Reproduction and Genetics 10/2013 Go to the issue

Assisted Reproduction Technologies

“Follicular HCG endometrium priming for IVF patients experiencing resisting thin endometrium. A proof of concept study”

Technological Innovations

The heat is on: room temperature affects laboratory equipment–an observational study

Assisted Reproduction Technologies

Thyroid stimulating hormone levels rise after assisted reproductive technology

Fertility Preservation

Is transplantation of cryopreserved ovarian tissue from patients with advanced-stage breast cancer safe? A pilot study

Editor’s Commentary

Networking necessities in the world of big science

Fertility Preservation

VEGF and bFGF increase survival of xenografted human ovarian tissue in an experimental rabbit model