Long-term co-culture of bovine granulosa cells with microvascular endothelial cells: effect on cell growth and cell death

Spanel‐Borowski, Katharina; Ricken, Albert; Saxer, Markus; Huber, Peter

doi:10.1016/0303-7207(94)90046-9

Cited by 15 publications

(7 citation statements)

References 33 publications

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“…Primary cultures of bovine granulosa cells were established as previously described [30] to undertake labeled exosomes uptake experiment. For this, granulosa cells isolated from bovine follicles were washed once in calcium-magnesium free PBS (CMF) followed by centrifugation at 500X g for 5 minutes and resuspended in 2 ml of RBC lysis buffer for 3-5 minutes to remove erythrocytes.…”

Section: Methodsmentioning

confidence: 99%

Exosomal and Non-Exosomal Transport of Extra-Cellular microRNAs in Follicular Fluid: Implications for Bovine Oocyte Developmental Competence

et al. 2013

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Cell-cell communication within the follicle involves many signaling molecules, and this process may be mediated by secretion and uptake of exosomes that contain several bioactive molecules including extra-cellular miRNAs. Follicular fluid and cells from individual follicles of cattle were grouped based on Brilliant Cresyl Blue (BCB) staining of the corresponding oocytes. Both Exoquick precipitation and differential ultracentrifugation were used to separate the exosome and non-exosomal fraction of follicular fluid. Following miRNA isolation from both fractions, the human miRCURY LNA™ Universal RT miRNA PCR array system was used to profile miRNA expression. This analysis found that miRNAs were present in both exosomal and non-exosomal fraction of bovine follicular fluid. We found 25 miRNAs differentially expressed (16 up and 9 down) in exosomes and 30 miRNAs differentially expressed (21 up and 9 down) in non-exosomal fraction of follicular fluid in comparison of BCB- versus BCB+ oocyte groups. Expression of selected miRNAs was detected in theca, granulosa and cumulus oocyte complex. To further explore the potential roles of these follicular fluid derived extra-cellular miRNAs, the potential target genes were predicted, and functional annotation and pathway analysis revealed most of these pathways are known regulators of follicular development and oocyte growth. In order to validate exosome mediated cell-cell communication within follicular microenvironment, we demonstrated uptake of exosomes and resulting increase of endogenous miRNA level and subsequent alteration of mRNA levels in follicular cells in vitro. This study demonstrates for the first time, the presence of exosome or non-exosome mediated transfer of miRNA in the bovine follicular fluid, and oocyte growth dependent variation in extra-cellular miRNA signatures in the follicular environment.

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Section: Methodsmentioning

confidence: 99%

Exosomal and Non-Exosomal Transport of Extra-Cellular microRNAs in Follicular Fluid: Implications for Bovine Oocyte Developmental Competence

et al. 2013

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“…Bovine granulosa cells were derived from cells harvested from bovine antral follicles and cultures estimated as described previously (Spanel-Borowski et al 1994b). Subconfluent primary granulosa cell cultures were used as positive controls of Kir2.1 and Kv1.4 mRNA.…”

Section: Isolation and Cultivation Of Mvecsmentioning

confidence: 99%

Voltage-dependent potassium channels in cytokeratin-positive and cytokeratin-negative microvascular endothelial cells of the corpus luteum

Richter

Tscheudschilsuren

Eschke

et al. 2002

Cell and Tissue Research

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We have recently described cytokeratin-positive (CK(+)) and cytokeratin-negative (CK(-)) microvascular endothelial cells (MVECs) in the bovine corpus luteum. The two phenotypes show a different expression and release of adhesion molecules and cytokines. Since secretion of mediators is specifically regulated by the electrophysiological membrane parameters, this report will compare voltage-dependent potassium (K(+)) channels in the two cell types cultured under the same conditions. CK(+) and CK(-) MVECs derived from the microvascular bed of one organ differ in their K(+)-channel characteristics. In CK(-) MVECs, an inwardly rectifying K(+) current was discovered, showing the characteristics of the Kir2.1. CK(+) MVECs displayed a voltage-dependent K(+) current that activates rapidly on depolarization and inactivates very slowly, and is associated with a member of the Kv family. The mRNA for Kir2.1 was identified by RT-PCR in CK(-) and CK(+) MVECs, but there was no evidence of Kv1.4 mRNA in either of them. The function of Kir2.1 in CK(-) MVECs might be induced during cultivation, whereas CK(+) MVECs appear to be more resistant to environmental conditions and do not express an inward current.

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“…Bovine granulosa cells were derived from cells harvested from bovine antral follicles and cultures estimated as described before [11]. Subconfluent primary granulosa cell cultures were used as positive controls of Kir2.1 and Kv1.4 mRNA.…”

Section: Isolation and Cultivation Of Retinal And Choroidal Mvecmentioning

confidence: 99%

Identification of Inwardly Rectifying Potassium Channels in Bovine Retinal and Choroidal Endothelial Cells

Eschke

Richter²,

Brylla³

et al. 2002

Ophthalmic Res

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Ion channels were studied using the whole-cell patch clamp technique in bovine retinal and choroidal microvascular endothelial cells (MVEC) cultured under the same conditions. The two types of MVEC expressed inward currents at hyperpolarizing voltage steps and showed small outward currents at depolarizing steps. The extrapolated reversal potentials of the inward currents were near to the potassium equilibrium potential. Cs⁺ and the K⁺ channel blocker TEA reduced the amplitudes of the currents indicating the selectivity and permeability for potassium. This was confirmed by changes of outside K⁺ concentration shifting the I-V curves to the right. RT-PCR studies revealed the presence of mRNA of Kir2.1, an inwardly rectifying K⁺ channel, in retinal and choroidal MVEC. The profile of the small outward currents is related to the Kv family but not identical with the Kv1.4 subtype.

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Long-term co-culture of bovine granulosa cells with microvascular endothelial cells: effect on cell growth and cell death

Cited by 15 publications

References 33 publications

Exosomal and Non-Exosomal Transport of Extra-Cellular microRNAs in Follicular Fluid: Implications for Bovine Oocyte Developmental Competence

Exosomal and Non-Exosomal Transport of Extra-Cellular microRNAs in Follicular Fluid: Implications for Bovine Oocyte Developmental Competence

Voltage-dependent potassium channels in cytokeratin-positive and cytokeratin-negative microvascular endothelial cells of the corpus luteum

Identification of Inwardly Rectifying Potassium Channels in Bovine Retinal and Choroidal Endothelial Cells

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