Secretory characteristics and viability of human term placental tissue after overnight cold preservation

Cirelli, Nadia; Lebrun, Philippe; Gueuning, C; Moens, André; Delogne-Desnoeck, J.; Dictus-Vermeulen, C.; Vanbellinghen, Anne-Marie; Meuris, Sylvain

doi:10.1093/humrep/15.4.756

Cited by 17 publications

(9 citation statements)

References 27 publications

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“…According to Cirelli et al ., 80% of LDH release occurs during the first 3 hours of incubation [29]. This supports our approach, assessing each fragment for 2 hours in the medium to quantify cytotoxicity after freezing…”

Section: Discussionsupporting

confidence: 76%

Examination of viability and quality of ovarian tissue after cryopreservation using simple laboratory methods in ewe

Merdassi

Mazoyer

Guérin

et al. 2011

Reprod Biol Endocrinol

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BackgroundThe objective of the present study is to assess viability tests and to evaluate follicle ovarian tissue quality after freezing-thawing procedures.MethodsEwe's ovaries were harvested at the slaughterhouse, after dissection each ovarian specimen was divided into two groups: fresh tissue (control group) and frozen tissue.In the first part of the study, the follicles viability was assessed by trypan blue staining, calcein AM/ethidium homodimer-1 staining (LIVE/DEAD viability/cytotoxicity kit, Molecular Probes) and morphology in the two groups. In the second part of the study the quality of the whole ovarian tissue was evaluated by the quantification of the release of lactate dehydrogenase measurement (Cytotoxicity Detection kit ROCHE), DNA fragmentation by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL) in primordial and primary follicles (ApopDETEK Kit system Enzo) and morphology in the two groups. 100 Follicles (primordial and primary) were counted on both fresh and frozen hemiovary to assess this various tests.ResultsOvarian follicle viability assessment was similar using trypan blue or calcein/ethidium staining. Follicles showed a decreased viability after freezing-thawing.After cryopreservation, a significant correlation between the percentage of normal follicles and viability rate was found using trypan blue (r = 0.82, p < 0.05) or calcein AM/ethidium homodimer-1 staining (r = 0.76, p < 0.05). Increased cytotoxicity showed by enhancement of LDH release was found after cryopreservation (21.60 +/- 1.1% vs 52.2 +/- 7.7%). A significant negative correlation between the percentage of morphologically normal follicles and cytotoxicity was observed. No significant difference in DNA fragmentation rate between frozen and control groups was found (26 ± 8.2% vs 38 ± 4.5%).ConclusionWe suggest the use of trypan blue staining for the histological assessment of viability, the use of LDH assay for the cytotoxicity assessement and finally the use of DNA fragmentation assessment to valid different freezing-thawing protocols.

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Section: Discussionsupporting

confidence: 76%

Examination of viability and quality of ovarian tissue after cryopreservation using simple laboratory methods in ewe

Merdassi

Mazoyer

Guérin

et al. 2011

Reprod Biol Endocrinol

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“…The RIN values observed compared very favorably to those of RIN values for human placental samples collected and stored under ideal conditions (Martin et al 2017). Metabolic changes can be detected within a few minutes following separation of the human placenta from their uterine blood supply (Cirelli et al 2000, Serkova et al 2003. These metabolic responses can include a halving or more of adenosine triphosphate concentrations rapidly (Serkova et al 2003).…”

Section: Discussionmentioning

confidence: 99%

Steroidogenic enzyme activities in the pre- and post-parturient equine placenta

Legacki¹,

Corbin²,

Ball³

et al. 2018

Reproduction

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Steroidogenic enzymes in placentas shape steroid hormone profiles in the maternal circulation of each mammalian species. These include 3β-hydroxysteroid dehydrogenase/Δ5-4 isomerase (3βHSD) and 17α-hydroxylase/17,20-lyase cytochrome P450 (P450c17) crucial for progesterone and androgen synthesis, respectively, as well as aromatase cytochrome P450 (P450arom) that converts Δ4-androgens to estrogens. 5α-reductase is another important enzyme in equine placentas because 5α-dihydroprogesterone (DHP) sustains pregnancy in the absence of progesterone in the second half of equine pregnancy. DHP and its metabolites decline dramatically days before foaling, but few studies have investigated placental enzyme activity before or at parturition in mares. Thus, key enzyme activities and transcript abundance were investigated in equine placentas at 300 days of gestation (GD300) and post-partum (term). Equine testis was used as a positive control for P450c17 activity. Substrates were incubated with microsomal preparations, together with enzyme inhibitors, and products were measured by liquid chromatography tandem mass spectrometry or radiometric methods (aromatase). Equine placenta expressed high levels of 3βHSD, 5α-reductase and aromatase, and minimal P450c17 activity at GD300 compared with testis (600-fold higher). At foaling, 3βHSD and aromatase activities and transcript abundance were unchanged but 5α-reductase (and P450c17) was no longer detectable ( < 0.05) and transcript was decreased. Trilostane inhibited 3βHSD significantly more in testis than placenta, suggesting possible existence of different 3βHSD isoforms. Equine placentas have significant capacity for steroid metabolism by 5α-reductase, 3βHSD and aromatase but little for androgen synthesis lacking P450c17. Declining pre-partum 5α-reduced pregnane concentrations coincide with selective loss of placental 5α-reductase activity and expression at parturition in horses.

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“…Caspases are well known for their role in cell suicide by apoptosis, but recent studies indicate they also play a role in cell proliferation and differentiation (10 -12). Knowledge of the expression and activity of caspases in placental villi and cultured trophoblasts is in its infancy (13)(14)(15). The proforms of the initiator caspase 8 and the effector caspases 3, 6, and 7 are expressed in villous cytotrophoblast and syncytiotrophoblasts.…”

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confidence: 99%

Trophoblast Differentiation Modulates the Activity of Caspases in Primary Cultures of Term Human Trophoblasts

Yusuf

2002

Pediatric Research

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Cultured human cytotrophoblasts are more susceptible than syncytiotrophoblasts to hypoxia-induced apoptosis. Caspases are cysteine proteases that cleave cellular components to effect the apoptotic cascade. We hypothesized that cultured cytotrophoblasts exhibit a higher activity of caspases when compared with syncytiotrophoblasts. Using western analysis, we demonstrated that the pro-caspases 3, 6, 8, and 9 are expressed in cytotrophoblasts cultured for 24 h, and also, in trophoblasts cultured 72 h when syncytiotrophoblasts have formed. Importantly, we found significantly higher activity of all four caspases in trophoblasts cultured 24 h compared with cells cultured 72 h. Colchicine and DMSO, which hinder trophoblast differentiation, enhanced the activity of all four caspases in cells cultured 72 h. Conversely, caspase activity was reduced in trophoblasts cultured for 24 h in the presence of epidermal growth factor, which enhances differentiation. This effect was most pronounced on caspase 3 and was attenuated by addition of the tyrosine kinase inhibitor AG1478. We conclude that cytotrophoblasts exhibit a higher activity of caspases 3, 6, 8, and 9 when compared with the more differentiated syncytium. This may account for the higher susceptibility of cytotrophoblasts to hypoxia-induced apoptosis. Human placental villi are covered by terminally differentiated, multinucleated syncytiotrophoblasts that are in direct contact with maternal blood. The subjacent mononucleated cytotrophoblasts provide a proliferative, stem cell population that differentiates and fuses to replenish the syncytium. Turnover of the trophoblast bi-layer occurs, in part, through apoptosis (1, 2). The balance of proliferation, differentiation, and apoptosis in the trophoblast layer of villi ultimately determines the mass of functional trophoblast available to regulate nutrient and waste exchange between the maternal and fetal circulations. Preeclampsia and fetal growth restriction (FGR) are associated with placental dysfunction, including villous hypoxia, altered differentiation and enhanced apoptosis in trophoblast (3-6). We previously showed that hypoxia hinders differentiation (7) and enhances apoptosis (8) in cultured trophoblasts from term placentas. Importantly, we found that the cytotrophoblasts were more susceptible than syncytiotrophoblasts to hypoxia-induced apoptosis.The regulation of apoptotic cell death is complex (9). Multiple ligand-receptor interactions and diverse stimuli that modulate mitochondrial function converge to enhance the activity of initiator caspases, cysteine proteases that self-amplify their enzymatic activity when stimulated. These enzymes activate downstream effector caspases that cleave membrane, cytoplasmic, and nuclear components, effectively dismantling the cell. Caspases are well known for their role in cell suicide by apoptosis, but recent studies indicate they also play a role in cell proliferation and differentiation (10 -12). Knowledge of the expression and activity of caspases in placental villi and cult...

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Secretory characteristics and viability of human term placental tissue after overnight cold preservation

Cited by 17 publications

References 27 publications

Examination of viability and quality of ovarian tissue after cryopreservation using simple laboratory methods in ewe

Examination of viability and quality of ovarian tissue after cryopreservation using simple laboratory methods in ewe

Steroidogenic enzyme activities in the pre- and post-parturient equine placenta

Trophoblast Differentiation Modulates the Activity of Caspases in Primary Cultures of Term Human Trophoblasts

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