Equine chorionic gonadotropin induces in vitro follicular growth from the multi-layered secondary developmental stage in cats

Chansaenroj, Ajjima; Songsasen, Nucharin; Chatdarong, Kaywalee

doi:10.1016/j.theriogenology.2018.09.040

Cited by 7 publications

(16 citation statements)

References 41 publications

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“…Oocyte diameters were similar in G‐0%, G‐0.5% and G‐1% follicles, in consistency with previous studies (Chansaenroj et al, 2019). In line with the observed increase in follicular diameters, all the groups showed increased oocyte diameters (approximately 75–76 μm, on day 4 of culture).…”

Section: Discussionsupporting

confidence: 92%

“…For this reason, it is essential to determine the adequate concentration of alginate that simultaneously maintains follicle structure during folliculogenesis in the feline species, providing support and permeability to the follicle, and thus ensures an adequate supply of nutrients, hormones and local regulatory factors towards the encapsulated follicle (Xu et al, 2009). In cat, preantral follicles have been cultured either in 2D (Jewgenow & Göritz, 1995; Wongbandue et al, 2013) or alginate‐based 3D culture systems with 0.3% (Nagashima et al, 2021) or 0.5% (Chansaenroj et al, 2019; Songsasen et al, 2012; Songsasen, Thongkittidilok, et al, 2017; Thongkittidilok et al, 2018) of this compound based on previous concentrations used in dogs (Songsasen, Nagashima, et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

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Determination of the appropriate concentration of sodium alginate used for in vitro culture of cat preantral follicles in a serum‐free medium containing FSH, EGF and IGF‐I

Fuertes-Recuero

González-Gil

Pérez

et al. 2023

Reprod Domestic Animals

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Culture of domestic cat preantral follicles can be a suitable technology to assist oocyte conservation strategies in the family Felidae. This research was aimed to comparatively analyse cat preantral follicular development of follicles directly seeded on growth surface or encapsulated in 0.5 or 1% of sodium alginate in a serum-free medium containing FSH, EGF and IGF-I. Preantral follicles were isolated from cat ovarian cortical tissue after ovariectomy. Alginate was dissolved at 0.5 or 1% in PBS. Follicles, 4 per well, with 0% (G-0%), 0.5% (G-0.5%) or 1% (G-1%) of sodium alginate were cultured in M199 with FSH (100 ng/mL), EGF (100 ng/mL) and IGF-I (100 ng/mL) for 7 days at 37°C, 5% CO 2 and 99% humidity. Culture medium was replaced every 48 h and samples were stored at −20°C until ELISA of steroid hormones. Morphometric evaluation of follicles was performed every 24 h. G-0% follicles showed granulosa cell migration away from the oocyte and disrupted morphology, whereby they reached apparently larger diameters (203.70 ± 5.82 μm; p < .05) than G-0.5% and G-1% follicles (157.89 ± 8.47 μm and 95.23 ± 1.67 μm, respectively) which maintained threedimensional organization, being larger in G-0.5% than in G-1% (p < .05). G-0.5% follicles attained the multi-layer preantral follicle stage on day 7 of culture, whereas G-1% follicles underwent progressive atresia. On day 6, steroid concentrations were higher (p < .

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Section: Discussionsupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Determination of the appropriate concentration of sodium alginate used for in vitro culture of cat preantral follicles in a serum‐free medium containing FSH, EGF and IGF‐I

Fuertes-Recuero

González-Gil

Pérez

et al. 2023

Reprod Domestic Animals

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“…cDNA was synthesized from 100 ng mRNA using a Transcriptor High Fidelity cDNA synthesis kit (Roche), according to the manufacturer’s instructions. Expression of the following genes was evaluated: β-actin (( Thongkittidilok et al 2018 ), a reference gene), aromatase ( CYP19A1 ( Thongkittidilok et al 2018 ), a measure of steroidogenesis in granulosa cells), growth differentiation factor 9 ( GDF9 ( Chansaenroj et al 2019 ), oocyte-derived factor necessary for theca cell layer development ( Dong et al 1996 )), luteinizing hormone receptor ( LHR , a marker for theca cell function), STAR protein ( ( Thongkittidilok et al. 2018 ) a regulator of cholesterol transfer for theca cell steroidogenesis), and vascular endothelial growth factor ( VEGFA , a potential marker of theca cell presence and functionality).…”

Section: Methodsmentioning

confidence: 99%

In vitro development of mechanically and enzymatically isolated cat ovarian follicles

Nagashima

Hill

Songsasen

2021

Reproduction and Fertility

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Isolation of ovarian follicles is a key step in culture systems for large mammalian species, to promote continued growth of follicles beyond the preantral stage in fertility preservation efforts. Still, mechanical isolation methods are user-skill dependent and time consuming, whereas enzymatic strategies carry increased risk of damaging theca cell layers and the basement membranes. Here, we sought to determine the optimal method to rescue domestic cat (Felis catus) early antral and antral stage follicles from ovarian tissue and evaluate the influence of isolation strategy on follicle development, survival, and gene expression during 14 day in vitro culture in alginate hydrogel. Mechanical isolation was compared with 90 min digestion in 0.7 and 1.4 Wünsch units/ml Liberase blendzyme (0.7L and 1.4L, respectively). Mechanical isolation resulted in improved follicle growth and survival, and better antral cavity and theca cell maintenance in vitro, compared with 1.4L (P<0.05), but displayed higher levels of apoptosis after incubation compared with enzymatically isolated follicles. However, differences in follicle growth and survival were not apparent until 7+ days in vitro. Expression of CYP19A1, GDF9, LHR, or VEGF were similar among isolation-strategies. Cultured follicles from all isolation methods displayed reduced StAR expression compared with freshly-isolated follicles obtained mechanically or via 0.7L, suggesting that prolonged culture resulted in loss of theca cell presence and/or function. In sum, early antral and antral stage follicle development in vitro is significantly influenced by isolation strategy, but not necessarily observable in the absence of extended culture. These results indicate additional care must be taken in follicle isolation optimizations for genome rescue and fertility preservation efforts.

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“…For this reason, we used planned comparisons for all analyses. We used a vehicle-control and FSH supplementation to determine whether FSH protected follicle viability, as expected based on previous work in this system (Songsasen et al, 2012; Chansaenroj, Songsasen & Chatdarong, 2019). We compared FSH-treatment alone to each concentration of fRFRP3 (1 and 10 µM) in combination with FSH to determine whether fRFRP3 has any effect on proposed outcome measures.…”

Section: Methodsmentioning

confidence: 99%

“…We predicted that fRFRP3 and its receptors, NPFFR1 and NPFFR2, would be expressed in the ovary. Second, we asked whether fRFRP3 impacts ovarian follicle integrity, survival, and steroid production by using an alginate-embedding system to culture isolated follicles (Songsasen et al, 2012; Thongkittidilok et al, 2018; Chansaenroj, Songsasen & Chatdarong, 2019). Based on studies in other mammals, we expected fRFRP3 to dose-dependently inhibit follicle growtth and to result in decreased viability of follicles (Maddineni et al, 2008; Wang, Li & Hu, 2018).…”

Section: Introductionmentioning

confidence: 99%

RFRP3 influences basal lamina degradation, cellular death, and progesterone secretion in cultured preantral ovarian follicles from the domestic cat

Wilsterman

Bentley

Comizzoli

2019

PeerJ

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The hypothalamic neuropeptide RFRP3 can suppress hypothalamic GnRH neuron activation and inhibit gonadotropin release from the anterior pituitary. RFRP3 is also produced locally in the ovary and can inhibit steroidogenesis and follicle development in many vertebrates. However, almost nothing is known about the presence and regulatory action of RFRP3 in gonads of any carnivore species. Such knowledge is important for developing captive breeding programs for endangered carnivores and for inhibiting reproduction in feral species. Using the domestic cat as a model, our objectives were to (1) demonstrate the expression of feline RFRP3 (fRFRP3) and its receptor in the cat ovary and (2) assess the influence of fRFRP3 on ovarian follicle integrity, survival, and steroidogenesis in vitro. We first confirmed that fRFRP3 and its receptors (NPFFR1 and NPFFR2) were expressed in cat ovaries by sequencing PCR products from ovarian RNA. We then isolated and cultured preantral ovarian follicles in the presence of 10 or 1 µM fRFRP3 + FSH (1 µg/mL). We recorded the percentage of morphologically viable follicles (basal lamina integrity) over 8 days and calculated percentage survival of follicles on Day 8 (using fluorescent markers for cell survival and death). Last, we quantified progesterone accumulation in media. 10 µM fRFRP3 had no observable effect on viability, survival, or steroid production compared to follicles exposed to only FSH. However, 1 µM fRFRP3 decreased the percentage of morphologically viable follicles and the percentage of surviving follicles on Day 8. At the same time, 1 µM fRFRP3 increased the accumulation of progesterone in media. Our study shows, for the first time, direct action of RFRP3 on the follicle as a functional unit, and it is the first in a carnivore species. More broadly, our results support a conserved, inhibitory action of RFRP3 on ovarian follicle development and underscore the importance of comparative functional studies.

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Equine chorionic gonadotropin induces in vitro follicular growth from the multi-layered secondary developmental stage in cats

Cited by 7 publications

References 41 publications

Determination of the appropriate concentration of sodium alginate used for in vitro culture of cat preantral follicles in a serum‐free medium containing FSH, EGF and IGF‐I

Determination of the appropriate concentration of sodium alginate used for in vitro culture of cat preantral follicles in a serum‐free medium containing FSH, EGF and IGF‐I

In vitro development of mechanically and enzymatically isolated cat ovarian follicles

RFRP3 influences basal lamina degradation, cellular death, and progesterone secretion in cultured preantral ovarian follicles from the domestic cat

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