scholarly journals Effects of In Vitro Interactions of Oviduct Epithelial Cells with Frozen–Thawed Stallion Spermatozoa on Their Motility, Viability and Capacitation Status

Animals ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 74
Author(s):  
Brenda Florencia Gimeno ◽  
María Victoria Bariani ◽  
Lucía Laiz-Quiroga ◽  
Eduardo Martínez-León ◽  
Micaela Von-Meyeren ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Sperm Quality ◽  
Assisted Reproduction Techniques ◽  
Cytokeratin Expression ◽  
Culture Model ◽  
Cryopreserved Sperm ◽  
Oviduct Epithelial Cells ◽  
Sperm Population ◽  
Selection Of

Cryopreservation by negatively affecting sperm quality decreases the efficiency of assisted reproduction techniques (ARTs). Thus, we first evaluated sperm motility at different conditions for the manipulation of equine cryopreserved spermatozoa. Higher motility was observed when spermatozoa were incubated for 30 min at 30 × 106/mL compared to lower concentrations (p < 0.05) and when a short centrifugation at 200× g was performed (p < 0.05). Moreover, because sperm suitable for oocyte fertilization is released from oviduct epithelial cells (OECs), in response to the capacitation process, we established an in vitro OEC culture model to select a sperm population with potential fertilizing capacity in this species. We demonstrated E-cadherin and cytokeratin expression in cultures of OECs obtained. When sperm–OEC cocultures were performed, the attached spermatozoa were motile and presented an intact acrosome, suggesting a selection by the oviductal model. When co-cultures were incubated in capacitating conditions a greater number of alive (p < 0.05), capacitated (p < 0.05), with progressive motility (p < 0.05) and with the intact acrosome sperm population was observed (p < 0.05) suggesting that the sperm population released from OECs in vitro presents potential fertilizing capacity. Improvements in handling and selection of cryopreserved sperm would improve efficiencies in ARTs allowing the use of a population of higher-quality sperm.

In vitro optimization of the Gallus domesticus oviduct epithelial cells culture

2012 ◽  
Vol 77 (9) ◽  
pp. 1834-1845 ◽  
Author(s):  
K. Kasperczyk ◽  
A. Bajek ◽  
R. Joachimiak ◽  
K. Walasik ◽  
A. Marszalek ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Gallus Domesticus ◽  
Oviduct Epithelial Cells

scholarly journals A Tissue-Engineered Tracheobronchial In Vitro Co-Culture Model for Determining Epithelial Toxicological and Inflammatory Responses

Biomedicines ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 631
Author(s):  
Luis Soriano ◽  
Tehreem Khalid ◽  
Fergal J. O'Brien ◽  
Cian O'Leary ◽  
Sally-Ann Cryan
Keyword(s):  
Epithelial Cells ◽  
Inflammatory Responses ◽  
Bronchial Epithelial Cells ◽  
Lung Fibroblasts ◽  
Drug Induced ◽  
Bronchial Epithelial ◽  
Culture Model ◽  
Epithelial Cultures

Translation of novel inhalable therapies for respiratory diseases is hampered due to the lack of in vitro cell models that reflect the complexity of native tissue, resulting in many novel drugs and formulations failing to progress beyond preclinical assessments. The development of physiologically-representative tracheobronchial tissue analogues has the potential to improve the translation of new treatments by more accurately reflecting in vivo respiratory pharmacological and toxicological responses. Herein, advanced tissue-engineered collagen hyaluronic acid bilayered scaffolds (CHyA-B) previously developed within our group were used to evaluate bacterial and drug-induced toxicity and inflammation for the first time. Calu-3 bronchial epithelial cells and Wi38 lung fibroblasts were grown on either CHyA-B scaffolds (3D) or Transwell® inserts (2D) under air liquid interface (ALI) conditions. Toxicological and inflammatory responses from epithelial monocultures and co-cultures grown in 2D or 3D were compared, using lipopolysaccharide (LPS) and bleomycin challenges to induce bacterial and drug responses in vitro. The 3D in vitro model exhibited significant epithelial barrier formation that was maintained upon introduction of co-culture conditions. Barrier integrity showed differential recovery in CHyA-B and Transwell® epithelial cultures. Basolateral secretion of pro-inflammatory cytokines to bacterial challenge was found to be higher from cells grown in 3D compared to 2D. In addition, higher cytotoxicity and increased basolateral levels of cytokines were detected when epithelial cultures grown in 3D were challenged with bleomycin. CHyA-B scaffolds support the growth and differentiation of bronchial epithelial cells in a 3D co-culture model with different transepithelial resistance in comparison to the same co-cultures grown on Transwell® inserts. Epithelial cultures in an extracellular matrix like environment show distinct responses in cytokine release and metabolic activity compared to 2D polarised models, which better mimic in vivo response to toxic and inflammatory stimuli offering an innovative in vitro platform for respiratory drug development.

scholarly journals Effects of Different Maturation Media on In Vitro Produced Bovine Embryos Co-Cultured with Bovine Oviduct Epithelial Cells or Cumulus Cells

1995 ◽  
Vol 12 (1) ◽  
pp. 9-13 ◽  
Author(s):  
C. Larocca ◽  
S. Kmaid ◽  
J. Calvo ◽  
J.E. Romano ◽  
M. Viqueira ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cumulus Cells ◽  
Bovine Embryos ◽  
Oviduct Epithelial Cells

scholarly journals In vitro Selection of Probiotics for Microbiota Modulation in Normal-Weight and Severely Obese Individuals: Focus on Gas Production and Interaction With Intestinal Epithelial Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Alicja Maria Nogacka ◽  
Clara G. de los Reyes-Gavilán ◽  
Silvia Arboleya ◽  
Patricia Ruas-Madiedo ◽  
Ceferino Martínez-Faedo ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Intestinal Epithelial Cells ◽  
Normal Weight ◽  
Gas Production ◽  
Intestinal Epithelial ◽  
Microbiota Composition ◽  
Severely Obese ◽  
Obese Subjects ◽  
Selection Of

The intestinal microbiota plays important roles in the maintenance of health. Strategies aiming at its modulation, such as probiotics, have received a deal of attention. Several strains have been studied in different in vitro models; however, the correlation of results obtained with the in vivo data has been limited. This questions the usefulness of such in vitro selection models, traditionally relying on over-simplified tests, not considering the influence of the accompanying microbiota or focusing on microbiota composition without considering functional traits. Here we assess the potential of six Bifidobacterium, Lactobacillus and Lacticaseibacillus strains in an in vitro model to determine their impact on the microbiota not just in terms of composition but also of functionality. Moreover, we compared the responses obtained in two different population groups: normal-weight and severely obese subjects. Fecal cultures were conducted to evaluate the impact of the strains on specific intestinal microbial groups, on the production of short-chain fatty acids, and on two functional responses: the production of gas and the interaction with human intestinal epithelial cells. The response to the different probiotics differed between both human groups. The addition of the probiotic strains did not induce major changes on the microbiota composition, with significant increases detected almost exclusively for the species added. Higher levels of gas production were observed in cultures from normal-weight subjects than in the obese population, with some strains being able to significantly reduce gas production in the latter group. Moreover, in obese subjects all the Bifidobacterium strains tested and Lacticaseibacillus rhamnosus GG were able to modify the response of the intestinal cells, restoring values similar to those obtained with the microbiotas of normal-weight subjects. Our results underline the need for the screening and selection of probiotics in a target-population specific manner by using appropriate in vitro models before enrolling in clinical intervention trials.

scholarly journals Modelling early events in Mycobacterium bovis infection using a co-culture model of the bovine alveolus

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Diane Frances Lee ◽  
Graham Roger Stewart ◽  
Mark Andrew Chambers
Keyword(s):  
Epithelial Cells ◽  
Conditioned Medium ◽  
Mycobacterium Bovis ◽  
Animal Health ◽  
Constitutive Expression ◽  
Alveolar Type ◽  
Cytokine Release ◽  
Peripheral Blood Mononuclear ◽  
Culture Model

Abstract Bovine tuberculosis (bTB), a zoonosis mainly caused by Mycobacterium bovis has severe socio-economic consequences and impact on animal health. Host–pathogen interactions during M. bovis infection are poorly understood, especially early events which are difficult to follow in vivo. This study describes the utilisation of an in vitro co-culture model, comprising immortalised bovine alveolar type II (BATII) epithelial cells and bovine pulmonary arterial endothelial cells (BPAECs). When cultured at air–liquid interface, it was possible to follow the migration of live M. bovis Bacille Calmette-Guérin (BCG) and to observe interactions with each cell type, alongside cytokine release. Infection with BCG was shown to exert a detrimental effect primarily upon epithelial cells, with corresponding increases in IL8, TNFα, IL22 and IL17a cytokine release, quantified by ELISA. BCG infection increased expression of CD54, MHC Class I and II molecules in endothelial but not epithelial cells, which exhibited constitutive expression. The effect of peripheral blood mononuclear cell conditioned medium from vaccinated cattle upon apical-basolateral migration of BCG was examined by quantifying recovered BCG from the apical, membrane and basolateral fractions over time. The numbers of recovered BCG in each fraction were unaffected by the presence of PBMC conditioned medium, with no observable differences between vaccinated and naïve animals.

Embryo co-culture with bovine amniotic membrane stem cells can enhance the cryo-survival of IVF-derived bovine blastocysts comparable with co-culture with bovine oviduct epithelial cells

Zygote ◽  
2020 ◽  
pp. 1-6
Author(s):  
Shayan Nejat-Dehkordi ◽  
Ebrahim Ahmadi ◽  
Abolfazl Shirazi ◽  
Hassan Nazari ◽  
Naser Shams-Esfandabadi
Keyword(s):  
Stem Cells ◽  
Growth Factors ◽  
Epithelial Cells ◽  
Embryonic Development ◽  
Amniotic Membrane ◽  
Biological Activities ◽  
Survival Rates ◽  
Blastocyst Stage ◽  
Oviduct Epithelial Cells

Summary Culture conditions have a profound effect on the quality of in vitro-produced embryos. Co-culturing embryos with somatic cells has some beneficial effects on embryonic development. Considering the ability of stem cells to secrete a broad range of growth factors with different biological activities, we hypothesized that bovine amniotic membrane stem cells (bAMSCs) might be superior to bovine oviduct epithelial cells (BOECs) in supporting embryonic development and enhancing their cryo-survival. Bovine abattoir-derived oocytes were matured and fertilized in vitro. The resultant presumptive zygotes were then cultured up to the blastocyst stage in the following groups: (i) co-culture with bAMSCs, (ii) co-culture with BOECs, and (iii) cell-free culture (Con). Embryos that reached the blastocyst stage were vitrified and warmed, and their post-warming re-expansion, survival and hatching rates were evaluated after 72 h culture. Results showed that the cleavage, blastocyst, and 2 h post-warming re-expansion rates of embryos did not differ between groups. However, their survival rates in BOEC and bAMSC groups were significantly higher compared with the control (72.7, 75.6 and 37.5%, respectively, P < 0.05). In conclusion, our results showed that the cryo-survivability of IVF-derived bovine embryos could be improved through co-culturing with bAMSCs. Moreover, considering the possibility to provide multiple passages from bAMSCs compared with BOECs, due to their stemness properties and their ability to produce growth factors, the use of bAMSCs is a good alternative to BOECs in embryo co-culture systems.

scholarly journals Influence of co-culture during maturation on the developmental potential of equine oocytes fertilized by intracytoplasmic sperm injection (ICSI)

Reproduction ◽  
2001 ◽  
pp. 925-932 ◽  
Author(s):  
X Li ◽  
LH Morris ◽  
WR Allen
Keyword(s):  
Epithelial Cells ◽  
Intracytoplasmic Sperm Injection ◽  
Fibroblast Cells ◽  
Cell Stage ◽  
Developmental Potential ◽  
Fetal Fibroblast ◽  
Nuclear Maturation ◽  
Fetal Fibroblasts ◽  
Oviduct Epithelial Cells

The influence of co-culture with either oviduct epithelial cells or fetal fibroblast cells on in vitro maturation of equine oocytes and their potential for development to blastocysts and fetuses after intracytoplasmic sperm injection (ICSI) was investigated. The oocytes were obtained from ovaries from abattoirs and were matured in vitro for 28-30 h in TCM-199 only, or in TCM-199 co-culture with oviduct epithelial cells or fetal fibroblast cells. Metaphase II oocytes were subjected to ICSI with an ionomycin-treated spermatozoon. The injected oocytes were cultured for 7-9 days in Dulbecco's modified Eagle's medium. Morphologically normal early blastocysts were transferred to the uteri of recipient mares. Nuclear maturation rates and the rates of cleavage to the two-cell stage for injected oocytes were similar in the groups of oocytes that were matured in TCM-199 (49 and 63%), in co-culture with oviduct epithelial cells (53 and 65%) or in co-culture with fetal fibroblasts (51 and 57%). There were no significant differences in the proportions of blastocysts that developed from the two-cell embryos derived from oocytes matured by co-culture with either oviduct epithelial cells (30%) or fetal fibroblasts (17%). However, significantly higher proportions of blastocysts were produced from both these co-culture groups than from the groups of oocytes matured in TCM-199 only (P < 0.05). Six of the blastocysts that had developed from oocytes co-cultured with oviduct epithelial cells were transferred into recipient mares and four pregnancies resulted. These results demonstrate a beneficial influence of co-culture with either oviduct epithelial cells or fetal fibroblasts for maturation of oocytes in vitro.

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