Culture of endometrial epithelial cells collected by a cytological brush in vivo

Rocha, Cecília Constantino; Silva, Felipe Alves Correa Carvalho; Martins, Thiago; Marrero, Marcela G.; Bromfield, John J.; Driver, John P.; Hansen, Thomas; Oliveira, Lilian J.; Binelli, Mário

doi:10.3168/jdsc.2021-0189

Cited by 5 publications

(3 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Here, we used a minimally invasive method (i.e., cytobrush) to allow recurrent sampling over time to probe uterine luminal epithelial cells [40]. These cells are fundamental for embryo–maternal signaling, as they serve as the first maternal layer of contact with the embryo, being responsible for the secretion and trafficking of molecules [71].…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Hormonal profile prior to luteolysis modulates the uterine luminal transcriptome in the subsequent cycle in beef cross-bred cows

Martins

Sponchiado

Rocha

et al. 2023

Biology of Reproduction

Self Cite

View full text Add to dashboard Cite

Sex steroid concentrations modulate endometrial function and fertility in cattle. Our objective was to compare the post-estrus luminal transcriptome of cows that were exposed to contrasting concentrations of progesterone (P4) before luteolysis that displayed estrus and ovulated spontaneously. Cross-bred beef cows received either 1) a new CIDR and GnRH (d − 9; high progesterone treatment; HP4; n = 16) or 2) a previously used CIDR, PGF2α, and GnRH (low progesterone treatment; LP4; n = 24). All cows received PGF2α at CIDR removal (d − 2). Ovarian ultrasonography and blood collections were performed on d − 9, d − 2, d − 0.5, d 0 (day of observed estrus), d 4, d 7, and d 14 for measurement of ovarian structures, P4, and estradiol (E2). Luminal epithelial cells were collected using a cytology brush on d 4, d 7, and d 14 for RNAseq. On d − 2, CL area and concentrations of P4 were greater, while on d − 0.5 concentrations of E2 were decreased in HP4. Ovarian structures and hormonal concentrations were similar on d 4, d 7, or d 14 (P > 0.05). There were enriched pathways in HP4 related to activation and signaling of the innate immune system at d 4, downregulation in the network involved in the extracellular matrix remodeling at d 7, and exacerbated inflammatory response as well as differentiation and activation of macrophages at d 14 (Benjamini – Hochberg P-value ≤0.05). In conclusion, manipulation of pre-luteolysis sex steroid concentrations altered the post-estrus luminal transcriptome even though all cows showed estrus and ovulated spontaneously.

show abstract

Section: Discussionmentioning

confidence: 99%

“…The brush was added back to the microcentrifuge tube containing the remaining cell pellet, snap-frozen, and stored at −80 • C for subsequent transcriptomic analysis. Cells obtained by this methodology were expected to be predominantly luminal epithelial cells [40].…”

Section: Cytobrush Collection and Processingmentioning

confidence: 99%

Hormonal profile prior to luteolysis modulates the uterine luminal transcriptome in the subsequent cycle in beef cross-bred cows

Martins

Sponchiado

Rocha

et al. 2023

Biology of Reproduction

Self Cite

View full text Add to dashboard Cite

show abstract

“…As a final point regarding the regulation of endometrial function: a lot less is known about sex-steroid-independent mechanisms of control. Martins and co-authors used the cytobrush to collect endometrial luminal epithelial cells (Rocha et al, 2022) four days after a synchronized estrus in crossbred beef embryo recipient cows (Martins et al, 2022). The authors compared the luminal epithelial transcriptome of cows that maintained vs. lost pregnancy after embryo transfer.…”

Section: The Multi-component Regulation Of Luminal Functionmentioning

confidence: 99%

Endometrial receptivity in cattle: the mutual reprogramming paradigm

et al. 2022

Self Cite

View full text Add to dashboard Cite

Prior to implantation in cattle, the mucous medium contained in the uterine lumen serves as a working interface for molecular exchange and signaling between the lining endometrium and the embryo. The composition of this luminal fluid changes temporally according to the secretory and reabsorptive activities of the uterus and the embryo, which are under complex regulation. Via this interface, both the embryo and the endometrium reprogram each other's functions to support pregnancy continuation beyond the pre-implantation period. More specifically, the embryo receives elongation signals and the uterus receives anti-luteolytic stimuli. Here, characteristics of the luminal compartment as well as the regulation of its composition to determine the pregnancy outcome will be discussed.

show abstract

In vitro reminiscence: uterine programming in vivo affects respective luminal epithelial cells function in vitro

Rocha,

Cordeiro,

Campbell

et al. 2024

Biology of Reproduction

View full text Add to dashboard Cite

In cattle, the endometrium during diestrus and early pregnancy displays cellular responses that are consequences of prior, transient stimuli. Goal was to establish a model to study cellular memory in the endometrium. The hypothesis is that stimuli given to endometrium in vivo are retained as a cellular memory that remains after bovine uterine epithelial cells (BUECs) are isolated, cultured, and further stimulated in vitro. Objectives were to measure BUEC proliferation/migration and responsiveness to recombinant bovine Interferon-tau (rbIFNT) in vitro: among cows that showed estrus (experiment 1 [Exp1]), cows that became or not pregnant to artificial insemination (Exp2), cows that received or not supplemental progesterone (P4; Exp3) and cows that received or not a COX-1/2 inhibitor (Exp4). Only cows that displayed estrus were included in studies. For all experiments endometrial cytology was collected 4 days after estrus, BUECs were cultured, propagated, and submitted to rbIFNT treatment and an in vitro scratch assay. In Exp1, different cows spontaneously grouped according to proliferative/migratory capacity and responsiveness to rbIFNT of their respective BUECs. In Exp2, BUECs from pregnant cows showed greater rbIFNT responsiveness and cellular proliferation. In Exp3, BUECs from cows supplemented with P4 presented inhibited proliferation and increased expression of RSAD2. In Exp4, Flunixin Meglumine modified rbIFNT responsiveness of BUECs in an IFN-signaling pathway-specific manner. In conclusion, physiological and pharmacological stimuli received by the endometrium in vivo were retained as cellular memory in BUECs, persisted in culture, and changed BUEC proliferation/migration and responsiveness to rbIFNT, which are characteristics associated with fertility in cattle.

show abstract

Culture of endometrial epithelial cells collected by a cytological brush in vivo

Cited by 5 publications

References 26 publications

Hormonal profile prior to luteolysis modulates the uterine luminal transcriptome in the subsequent cycle in beef cross-bred cows

Hormonal profile prior to luteolysis modulates the uterine luminal transcriptome in the subsequent cycle in beef cross-bred cows

Endometrial receptivity in cattle: the mutual reprogramming paradigm

In vitro reminiscence: uterine programming in vivo affects respective luminal epithelial cells function in vitro

Contact Info

Product

Resources

About