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Coenzyme Q₁₀ in Submicron-Sized Dispersion Improves Development, Hatching, Cell Proliferation, and Adenosine Triphosphate Content of In Vitro-Produced Bovine Embryos¹

^a Lehrstuhl für Molekulare Tierzucht und Haustiergenetik, Ludwig-Maximilians-Universität München, 85764 Oberschleissheim, Germany ^b Lehrstuhl für Pharmazeutische Technologie, Institut für Pharmazie, Friedrich-Schiller-Universität, 07743 Jena, Germany ^c Bayerisches Forschungszentrum für Fortpflanzungsbiologie GmbH & Co KG (BFZF), 85764 Oberschleissheim, Germany

Coenzyme Q₁₀ (CoQ₁₀) is an essential component of the plasma membrane ion transporter (PMIT) system and of the electron transport chain in the inner mitochondrial membrane. Because of its intrinsic functions in cell growth and energy metabolism (ATP synthesis), and its protective effects against oxidative stress, CoQ₁₀ is a good candidate for supporting growth of cells in culture. However, because of its quinone structure, CoQ₁₀ is extremely lipophilic and practically insoluble in water. We used a specific technology to prepare a submicron-sized dispersion of CoQ₁₀, inhibiting re-crystallization by a stabilizer. This dispersion, which exhibits a very large specific surface area for drug dissolution, was tested as a supplement for the in vitro culture of bovine embryos in a chemically defined system. The rate of early cleavage of embryos (5- to 8-cell stages) was evaluated 66 h postinsemination (hpi) and was highest in medium supplemented with 30 or 100 µM CoQ₁₀ (66.5 ± 0.8% and 68.7 ± 1.1%, respectively) and lowest in 10 µM CoQ₁₀ (55.3 ± 0.8%). The proportions of oocytes developing to blastocysts by 186 hpi were 19.0 ± 0.6% and 25.2 ± 0.3% in medium supplemented with 10 µM and 30 µM CoQ₁₀, respectively, and were significantly (p < 0.001) higher than those obtained with the equivalent amounts of stabilizer (9.9 ± 0.4% and 11.3 ± 0.4%). In the presence of 30 µM CoQ₁₀, significantly (p < 0.001) more blastocysts hatched by 210 hpi than in the equivalent amount of stabilizer (31.8 ± 1.3 vs. 8.4 ± 2.2). Expanded blastocysts produced in the presence of 30 µM CoQ₁₀ had significantly (p < 0.01) more inner cell mass cells and trophectoderm cells, and a significantly (p < 0.001) increased ATP content as compared to expanded blastocysts produced in the presence of the corresponding amount of stabilizer.

Our results show that noncrystalline CoQ₁₀ in submicron-sized dispersion supports the development and viability of bovine embryos produced in a chemically defined culture system.

¹ This study was supported by the Bayerische Forschungsstiftung (76/93) and by the DFG-WO 685/2-1; WO 685/3-1.

² Correspondence: Eckhard Wolf, Lehrstuhl für Molekulare Tierzucht und Haustiergenetik, Hackerstr. 27, 85764 Oberschleissheim, Germany. FAX: 49 89 74017 368; ewolf{at}lmb.uni-muenchen.de

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