HUMAN FOLLICLE STIMULATING HORMONE (hFSH) AND THYROXINE (T<sub>4</sub>)
               IN SURVIVAL MAINTENANCE AND IN VITRO GROWTH PROMOTION OF CAPRINE PREANTRAL
               FOLLICLES

Costa, Sanely Lourenço da; Costa, Eduardo Paulino da; Pereira, Emílio César Martins; Gonçalves, Wagner Gonzaga; Silva, Talita Fernandes da; Queiroz, Vanessa Lopes Dias

doi:10.1590/1089-6891v16i231471

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MEDICINA VETERINÁRIA • Ciênc. anim. bras. 16 (2) • Apr-Jun 2015 • https://doi.org/10.1590/1089-6891v16i231471 copy

HUMAN FOLLICLE STIMULATING HORMONE (hFSH) AND THYROXINE (T₄) IN SURVIVAL MAINTENANCE AND IN VITRO GROWTH PROMOTION OF CAPRINE PREANTRAL FOLLICLES

HORMÔNIO FOLÍCULO ESTIMULANTE HUMANO (hFSH) E TIROXINA (T₄) NA MANUTENÇÃO DA SOBREVIVÊNCIA E PROMOÇÃO DO CRESCIMENTO IN VITRO DE FOLÍCULOS PRÉ-ANTRAIS CAPRINOS

Authorship SCIMAGO INSTITUTIONS RANKINGS

The aim of this study was to investigate the interaction of human FSH (10ng/ml) with T₄ (20ng/mL) on survival, activation and growth of preantral follicles cultured in vitro for 28 days. Fragments of non-cultured and cultured ovarian tissue were processed for classic histology and transmission electron microscopy. The results showed a reduction in the survival rate in all the media tested (one to 28 days) when compared to the fresh control. However the treatment with T₄/hFSH for seven days of culture maintained the rate similar to the control. The media tested by one and 28 days reduced the percentage of primordial follicles in all periods of culture. However, T₄/hFSH on day one of culture remained similar to the fresh control. None of the media were able to keep the percentage of the developing follicles. It was observed that the follicular diameter in the medium with T₄/hFSH remained similar to the fresh control. The ultrastructural analysis confirmed the integrity of follicles cultured for seven days in a medium supplemented with T₄/hFSH. In conclusion, the medium with T₄/hFSH is able to maintain the survival, promote the activation, and the ultrastructural integrity of caprine preantral follicles for until seven days.

Activation; caprine; hFSH; preantral follicles; T₄

Culture medium	Abbreviation
α-MEM⁺(culture control for 1, 7, 14, 21 or 28 days)	α-MEM⁺
α-MEM⁺(T₄ + hFSH for 1, 7, 14, 21 or 28 days)	T₄/hFSH

Treatments
Control	97,3 ± 1,1
Control	Day l	Day 7	Day 14	Day 21	Day 28
α-MEM⁺	83.3 ± 2.0^* * Differs significantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	82.7 ± 1.9^* * Differs significantly from control follicles (P<0.05). ^b a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	78.0 ± 2.6^* * Differs significantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05). ^B A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	80.7 ± 1.6^* * Differs significantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	66.7 ± 4.6^* * Differs significantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^B A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).
T₄₉₍₂₀₎/hFSH₍₁₀₎	90.0 ± 0.0^* * Differs significantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	90.7 ± 1.6^b a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	80.7 ± 4.5^* * Differs significantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05). ^B A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	72.7 ± 2.6^* * Differs significantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05). ^B A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	66.0 ± 6.2^* * Differs significantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^B A,B differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).

Universidade Federal de Goiás Universidade Federal de Goiás, Escola de Veterinária e Zootecnia, Campus II, Caixa Postal 131, CEP: 74001-970, Tel.: (55 62) 3521-1568, Fax: (55 62) 3521-1566 - Goiânia - GO - Brazil
E-mail: revistacab@gmail.com

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[TFN01] ^*
Differs significantly from control follicles (P<0.05).

[TFN02] ^a,b
differs significantly among concentrations in each day of culture (P<0.05).

[TFN03] ^A,B
differs significautly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).

Primordial follicles
Treatments
Control	71.3 ± 4.2
Control	Day l	Day 7	Day 14	Day 21	Day 18
α-MEM⁺	42.7 ± 5.9^* * Differs from control follicles (P<0.05). ^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	34.7 ± 3.9^* * Differs from control follicles (P<0.05). ^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	42.0 ± 3.9^* * Differs from control follicles (P<0.05). ^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	32.0 ± 2.6^* * Differs from control follicles (P<0.05). ^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	38.0 ± 3.3^* * Differs from control follicles (P<0.05). ^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).
T₄₍₂₀₎/hFSH₍₁₀₎	48.7 ± 6.1^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	38.0 ± 8.1^* * Differs from control follicles (P<0.05). ^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	44.7 ± 8.4^* * Differs from control follicles (P<0.05). ^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	38.0 ± 4.4^* * Differs from control follicles (P<0.05). ^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	33.3 ± 7.9^* * Differs from control follicles (P<0.05). ^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).
Developing follicles
Treatments
Control	31.3 ± 7.4
Control	Day l	Day 7	Day 14	Day 21	Day 18
α-MEM⁺	40.7 ±7.1^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	48.0 ± 3.8^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	36.0 ± 4.7^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	48.7 ± 3.7^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	28.7 ± 3.2^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).
T₄₍₂₀₎/hFSH₍₁₀₎	41 3 ± 6.1^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	52.7 ± 9.4^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	36.0 ± 8.1^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	34.7 ± 3.3^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	32.7 ± 8.6^a a,b differs among concentrations in each dary of culture (P<0.05). ^A A,B differs with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).

Oocyte diameter (μm)
Treatments
Control	26.0 ± 0.6
Control	Day l	Day 7	Day 14	Day 21	Day 18
α-MEM⁺	24.2 ± 0.8^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	22.7 ± 0.7^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	21.2 ± 0.6^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	20.8 ± 1.0^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	20.3 ± 0.5^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).
T₄₍₂₀₎/hFSH₍₁₀₎	24.7 ± 0.8^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	23.7 ± 0.6^* * Differs signficantly from control follicles (P<0.05). ^A A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	20.7 ± 0.8^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05). ^C	21.7 ± 0.9^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05). ^C	19.6 ± 0.6^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^C
Follicle diameter (μm)
Treatments
Control	33.4 ± 0.8
Control	Day l	Day 7	Day 14	Day 21	Day 18
α-MEM⁺	32.5 ± 1.2^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	30.2 ± 1.2^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	27.8 ± 0.7^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	27.8 ± 1.0^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	27.e ± 0.7^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).
T₄₍₂₀₎/hFSH₍₁₀₎	33.2 ± 1.0^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	30.7 ± 0.7^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^A A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	27.5 ± 0.9^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05). ^C	29.0 ± 1.1.^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^B A,B differs significantly with the progression of the culture period from days 1 to 28 in the same treatment (P<0.05).	25.5 ± 0.6^* * Differs signficantly from control follicles (P<0.05). ^a a,b differs significantly among concentrations in each day of culture (P<0.05). ^C

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HUMAN FOLLICLE STIMULATING HORMONE (hFSH) AND THYROXINE (T4) IN SURVIVAL MAINTENANCE AND IN VITRO GROWTH PROMOTION OF CAPRINE PREANTRAL FOLLICLES

HORMÔNIO FOLÍCULO ESTIMULANTE HUMANO (hFSH) E TIROXINA (T4) NA MANUTENÇÃO DA SOBREVIVÊNCIA E PROMOÇÃO DO CRESCIMENTO IN VITRO DE FOLÍCULOS PRÉ-ANTRAIS CAPRINOS

HUMAN FOLLICLE STIMULATING HORMONE (hFSH) AND THYROXINE (T₄) IN SURVIVAL MAINTENANCE AND IN VITRO GROWTH PROMOTION OF CAPRINE PREANTRAL FOLLICLES

HORMÔNIO FOLÍCULO ESTIMULANTE HUMANO (hFSH) E TIROXINA (T₄) NA MANUTENÇÃO DA SOBREVIVÊNCIA E PROMOÇÃO DO CRESCIMENTO IN VITRO DE FOLÍCULOS PRÉ-ANTRAIS CAPRINOS