Preprint Article Version 1 This version is not peer-reviewed

In Vitro Bovine Embryo Production Using Medium’s Supplemented with Recombinant Human Galectin-1 Buffered

Version 1 : Received: 2 May 2022 / Approved: 13 May 2022 / Online: 13 May 2022 (15:14:42 CEST)

How to cite: Roncoletta, M.; Morani, E.D.S.C.; De Salles, N.A.A.; Ferraz, G.C. In Vitro Bovine Embryo Production Using Medium’s Supplemented with Recombinant Human Galectin-1 Buffered. Preprints 2022, 2022050193 (doi: 10.20944/preprints202205.0193.v1). Roncoletta, M.; Morani, E.D.S.C.; De Salles, N.A.A.; Ferraz, G.C. In Vitro Bovine Embryo Production Using Medium’s Supplemented with Recombinant Human Galectin-1 Buffered. Preprints 2022, 2022050193 (doi: 10.20944/preprints202205.0193.v1).

Abstract

As recommended in the ICH Guidelines (S5-R2 and S6-R1), and based on bioethical concerns, we chose bovine embryos (BE) to check the in vitro embryo development considering the use of different amounts of rHGAL-1 as supplementations of in vitro embryo culture (IVP) mediums. Based on procedures for commercial BE in vitro production, using oocytes aspirated from slaughterhouse ovaries, the rHGAL-1 supplementation performed in two experiments (#01 on the oocyte maturation - IVM medium supplemented and experiment #2 on culture step IVC, supplemented SOF medium). There were IVP commercial procedures done, with 3 IVP batches per experiment and distributed the oocytes in four groups of treatment (one control group and three different dosages of rHGAL-1 to supplement both IVM and SOF mediums, using (2, 20 and 40µg.mL-1 respectively). A total of 962 (experiment 1) and 1,213 (experiment 2) oocytes were aspirated and submitted to IVP procedure. There was no damage to in vitro bovine embryos growth, considering cleavage percentage (%CLE), blastocysts development on day 7 (BlD7, BxD7, BhD7), or hatching blastocysts maturation on day 8 (BhD8%), regardless of rHGAL-1 supplementation. The immunohistochemistry assay with D8 embryos cultivated with rHGAL-1 supplementation on the culture medium (SOF medium) could demonstrate the presence of exogenous GAL-1, distributed in mass cell and trophoblastic cells, and the profile observed is dependent of exogenous supplementation and it was more evident in hatched embryos. The findings reassure the use of a reasonable amount of rHGAL-1 for in vitro embryonic development and make using rHGAL-1 in assisted reproduction in humans more reliable and safer.

Keywords

Tolerana®; Pregnancy; Fetal-Maternal Recognition; Reproduction; Reproductive Safety Toxicology

Subject

MEDICINE & PHARMACOLOGY, Pharmacology & Toxicology

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