CHANGES IN THE CAPACITY FOR CLONAL GROWTH AND DIFFERENTIATION IN VITRO OF THE VERTEBRAL CARTILAGE CELLS WITH EMBRYONIC DEVELOPMENT. 1. CULTURE IN THE STANDARD MEDIUM

AbstractIn vitro culture can alter the development and quality of bovine embryos. Therefore, we aimed to evaluate whether nobiletin supplementation during EGA improves embryonic development and blastocyst quality and if it affects PI3K/AKT signaling pathway. In vitro zygotes were cultured in SOF + 5% FCS (Control) or supplemented with 5, 10 or 25 µM nobiletin (Nob5, Nob10, Nob25) or with 0.03% dimethyl-sulfoxide (CDMSO) during minor (2 to 8-cell stage; MNEGA) or major (8 to 16-cell stage; MJEGA) EGA phase. Blastocyst yield on Day 8 was higher in Nob5 (42.7 ± 1.0%) and Nob10 (44.4 ± 1.3%) for MNEGA phase and in Nob10 (61.0 ± 0.8%) for MJEGA phase compared to other groups. Mitochondrial activity was higher and lipid content was reduced in blastocysts produced with nobiletin, irrespective of EGA phase. The mRNA abundance of CDK2, H3-3B, H3-3A, GPX1, NFE2L2 and PPARα transcripts was increased in 8-cells, 16-cells and blastocysts from nobiletin groups. Immunofluorescence analysis revealed immunoreactive proteins for p-AKT forms (Thr308 and Ser473) in bovine blastocysts produced with nobiletin. In conclusion, nobiletin supplementation during EGA has a positive effect on preimplantation bovine embryonic development in vitro and corroborates on the quality improvement of the produced blastocysts which could be modulated by the activation of AKT signaling pathway.

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N-acetyl-L-cysteine Improves the Developmental Competence of Bovine Oocytes and Embryos Cultured In Vitro by Attenuating Oxidative Damage and Apoptosis

Antioxidants ◽

10.3390/antiox10060860 ◽

2021 ◽

Vol 10 (6) ◽

pp. 860

Author(s):

Wu-Sheng Sun ◽

Hoon Jang ◽

Mi-Ryung Park ◽

Keon Bong Oh ◽

Haesun Lee ◽

...

Keyword(s):

Embryonic Development ◽

Early Stage ◽

Developmental Competence ◽

Beneficial Effects ◽

Developmental Rates ◽

A Cell ◽

Bovine Oocytes ◽

Dose Dependent

Oxidative stress has been suggested to negatively affect oocyte and embryo quality and developmental competence, resulting in failure to reach full term. In this study, we investigated the effect of N-acetyl-L-cysteine (NAC), a cell-permeating antioxidant, on developmental competence and the quality of oocytes and embryos upon supplementation (0.1–10 mM) in maturation and culture medium in vitro using slaughterhouse-derived oocytes and embryos. The results show that treating oocytes with 1.0 mM NAC for 8 h during in vitro maturation attenuated the intracellular reactive oxygen species (ROS) (p < 0.05) and upregulated intracellular glutathione levels (p < 0.01) in oocytes. Interestingly, we found that NAC affects early embryonic development, not only in a dose-dependent, but also in a stage-specific, manner. Significantly (p < 0.05) decreased cleavage rates (90.25% vs. 81.46%) were observed during the early stage (days 0–2), while significantly (p < 0.05) increased developmental rates (38.20% vs. 44.46%) were observed during the later stage (from day 3) of embryonic development. In particular, NAC supplementation decreased the proportion of apoptotic blastomeres significantly (p < 0.05), resulting in enhanced hatching capability and developmental rates during the in vitro culture of embryos. Taken together, our results suggest that NAC supplementation has beneficial effects on bovine oocytes and embryos through the prevention of apoptosis and the elimination of oxygen free radicals during maturation and culture in vitro.

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Establishment of a New Device for Electrical Stimulation of Non-Degenerative Cartilage Cells In Vitro

International Journal of Molecular Sciences ◽

10.3390/ijms22010394 ◽

2021 ◽

Vol 22 (1) ◽

pp. 394

Author(s):

Simone Krueger ◽

Alexander Riess ◽

Anika Jonitz-Heincke ◽

Alina Weizel ◽

Anika Seyfarth ◽

...

Keyword(s):

Electrical Stimulation ◽

Electric Fields ◽

Cartilage Tissue ◽

Type I ◽

Dependent Manner ◽

New Device ◽

Alternating Electric Fields ◽

Cartilage Cells ◽

Stimulation Of

In cell-based therapies for cartilage lesions, the main problem is still the formation of fibrous cartilage, caused by underlying de-differentiation processes ex vivo. Biophysical stimulation is a promising approach to optimize cell-based procedures and to adapt them more closely to physiological conditions. The occurrence of mechano-electrical transduction phenomena within cartilage tissue is physiological and based on streaming and diffusion potentials. The application of exogenous electric fields can be used to mimic endogenous fields and, thus, support the differentiation of chondrocytes in vitro. For this purpose, we have developed a new device for electrical stimulation of chondrocytes, which operates on the basis of capacitive coupling of alternating electric fields. The reusable and sterilizable stimulation device allows the simultaneous use of 12 cavities with independently applicable fields using only one main supply. The first parameter settings for the stimulation of human non-degenerative chondrocytes, seeded on collagen type I elastin-based scaffolds, were derived from numerical electric field simulations. Our first results suggest that applied alternating electric fields induce chondrogenic re-differentiation at the gene and especially at the protein level of human de-differentiated chondrocytes in a frequency-dependent manner. In future studies, further parameter optimizations will be performed to improve the differentiation capacity of human cartilage cells.

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Effects of different concentrations of Chir98014 as an activator of Wnt/beta‐catenin signaling pathway on oocyte in‐vitro maturation and subsequent embryonic development in Sanjabi ewes

Reproduction in Domestic Animals ◽

10.1111/rda.13938 ◽

2021 ◽

Author(s):

Sarah Samereh ◽

Hadi Hajarian ◽

Hamed Karamishabankareh ◽

Leila Soltani ◽

Saheb Foroutanifar

Keyword(s):

Embryonic Development ◽

Signaling Pathway ◽

In Vitro Maturation ◽

Beta Catenin

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Effect of leptin during in vitro maturation of prepubertal calf oocytes: Embryonic development and relative mRNA abundances of genes involved in apoptosis and oocyte competence

Theriogenology ◽

10.1016/j.theriogenology.2011.07.002 ◽

2011 ◽

Vol 76 (9) ◽

pp. 1706-1715 ◽

Cited By ~ 11

Author(s):

Bladimir Córdova ◽

Roser Morató ◽

Celia de Frutos ◽

Pablo Bermejo-Álvarez ◽

Teresa Paramio ◽

...

Keyword(s):

Embryonic Development ◽

In Vitro Maturation ◽

Oocyte Competence ◽

Calf Oocytes

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Effect of D-Glucuronic Acid and N-acetyl-D-Glucosamine Treatment during In Vitro Maturation on Embryonic Development after Parthenogenesis and Somatic Cell Nuclear Transfer in Pigs

Animals ◽

10.3390/ani11041034 ◽

2021 ◽

Vol 11 (4) ◽

pp. 1034

Author(s):

Joohyeong Lee ◽

Eunhye Kim ◽

Seon-Ung Hwang ◽

Lian Cai ◽

Mirae Kim ◽

...

Keyword(s):

Embryonic Development ◽

Somatic Cell ◽

Somatic Cell Nuclear Transfer ◽

Nuclear Transfer ◽

In Vitro Maturation ◽

Glucuronic Acid ◽

Cumulus Cell ◽

Cortical Granule ◽

Oocyte Diameter

This study aimed to examine the effects of treatment with glucuronic acid (GA) and N-acetyl-D-glucosamine (AG), which are components of hyaluronic acid (HA), during porcine oocyte in vitro maturation (IVM). We measured the diameter of the oocyte, the thickness of the perivitelline space (PVS), the reactive oxygen species (ROS) level, and the expression of cumulus cell expansion and ROS-related genes and examined the cortical granule (CG) reaction of oocytes. The addition of 0.05 mM GA and 0.05 mM AG during the first 22 h of oocyte IVM significantly increased oocyte diameter and PVS size compared with the control (non-treatment). The addition of GA and AG reduced the intra-oocyte ROS content and improved the CG of the oocyte. GA and AG treatment increased the expression of CD44 and CX43 in cumulus cells and PRDX1 and TXN2 in oocytes. In both the chemically defined and the complex medium (Medium-199 + porcine follicular fluid), oocytes derived from the GA and AG treatments presented significantly higher blastocyst rates than the control after parthenogenesis (PA) and somatic cell nuclear transfer (SCNT). In conclusion, the addition of GA and AG during IVM in pig oocytes has beneficial effects on oocyte IVM and early embryonic development after PA and SCNT.

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Vitellogenin receptor selectively endocytoses female-specific and highly-expressed hemolymph proteins in the silkworm, Bombyx mori

Biochemistry and Cell Biology ◽

10.1139/bcb-2016-0255 ◽

2017 ◽

Vol 95 (4) ◽

pp. 510-516 ◽

Cited By ~ 10

Author(s):

Chaoshan Han ◽

Enxiang Chen ◽

Guanwang Shen ◽

Zhixin Peng ◽

Yinying Xu ◽

...

Keyword(s):

Embryonic Development ◽

Bombyx Mori ◽

Sf9 Cells ◽

Tissue Localization ◽

Vitellogenin Receptor ◽

Hemolymph Proteins ◽

Female Specific ◽

Oocyte Membrane ◽

Silkworm Bombyx Mori

VgR, a member of the LDLR family, functions to transport vitellogenin into the ovaries to protome ovarian growth and embryonic development. In insects, the only widely accepted ligand of VgR is Vg. Recently, BmVgR has been shown to interact with BmSP1 in vitro. Therefore, in this study, we evaluated whether BmVgR could transport BmSP1 into certain cells. Although BmVgR could combine with BmVg and BmSP1, BmVgR did not affect the amount of BmSP1 taken up by Sf9 cells. Parallel immunofluorescence showed that most BmVg and BmVgR were localized in the inner oocyte membrane, showing tissue localization similar to that of BmVg labeled with pHrodo Red absorbed by the ovaries on day 2 of pupation. Although BmSP1 showed localization similar to BmVgR during the same phase, little BmSP1 was present in the ovary. Additionally, BmSP1 did not exist in ovaries when the ovaries contained BmVgR on day 5 of pupation, suggesting that BmSP1 in the ovaries was not endocytosed by BmVgR. In summary, BmVgR could facilitate uptake of BmVg by developing oocytes, but did not modulate in the transport of BmSP1.

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