Effects of isolation and culture of turkey primary follicular oocytes on morphology and germinal vesicle integrity

Bakst, M. R.; Gliedt, D.W.; Akuffo, V.; Potts, William John; Gupta, Sandeep

doi:10.1016/s0093-691x(98)00213-1

Cited by 3 publications

(3 citation statements)

References 12 publications

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“…Trypsin, a proteolytic enzyme highly effective in cell disaggregation [22], and collagenase, which is capable of attacking native collagen without having an effect on related proteins and without damaging epithelial tissue, have been commonly used for isolating follicles from mammals [12]. However, using trypsin alone for dissection of the ovary as described by Bakst et al [18], the number of primary follicles (about 180) isolated from the hens were not satisfactory in this experiment. Furthermore, use of collagenase alone was also attempted to use in our laboratory, and this was also incapable of producing sufficient quantities of primary f oll icles f or further experiments.…”

Section: Discussionmentioning

confidence: 86%

“…Good follicles had a complete single layer of granulosa cells and a basement membrane. Follicles were classified as degenerated if they presented one of the following aspects: a shrunken oocyte, asymmetrical cytoplast, fully or partially denuded of granulosa cells, or no basement membrane [18,19].…”

Section: Classification Of Primary Follicles and Assessment Of Follicmentioning

confidence: 99%

“…Such techniques that have been suitable for isolation of mammal preantral follicles are not suitable for isolation of avian follicles. In 1998, 200 to 500 primary follicles were isolated from a turkey ovary using trypsin after the ovarian cortex was incubated for 30 to 120 min [18]. Following this success, an attempt was made to isolate primary follicles from the hen ovary in our laboratory.…”

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confidence: 99%

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An Efficient Isolation Method for Domestic Hen (Gallus domesticus) Ovarian Primary Follicles

Han

Jiang

et al. 2006

Journal of Reproduction and Development

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Abstract. An enzymatic method of isolating primary follicles in the turkey has been described previously, but no similar work has been done in the hen. In this study, primary follicles from domestic hens (Gallus domesticus) were isolated using an enzymatic method, and the isolated follicular quantity, quality, and development in vitro were assessed. About 400 primary follicles ranging from 60 to 1125 µm in diameter were recovered with trypsin and collagenase from hen ovaries (per ovary).Of these, 76.5% were intact follicles with a complete single layer of granulose cells surrounded by the basement membrane, and their ultrastructures appeared this way in situ. Follicles (351 to 500 µm in diameter) were cultured in vitro, and 46.67% of them survived after 5 days. Ultrastructural examination showed that elongated mitochondria forming a ring were distributed to the periphery of the oocyte, the Golgi was oriented with the maturing face toward the granulosa cell layer, and the oocyte plasma membrane presented a few short microvilli lying on the oocyte surface, which confirmed that the surviving follicles were developmental. These results suggest that a simple, rapid, effective enzymatic method can be used to isolate a great number of intact primary follicles from the hen ovary.

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Section: Discussionmentioning

confidence: 86%

Section: Classification Of Primary Follicles and Assessment Of Follicmentioning

confidence: 99%

mentioning

confidence: 99%

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An Efficient Isolation Method for Domestic Hen (Gallus domesticus) Ovarian Primary Follicles

Han

Jiang

et al. 2006

Journal of Reproduction and Development

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Development-promoting effect of chicken embryo membrane on chicken ovarian cortical pieces of different age

et al. 2009

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Ovarian cortical tissues of various ages of chicken were grafted underneath chorioallantoic membrane (CAM) of 6-d-old chicken embryo and the grafts were collected on d 0, 2, 4, 8, or 10. The tissue sections were prepared to examine the development of follicles in grafts and the proliferative ability of follicles was examined by immunohistochemistry analysis of proliferating cell nuclear antigen. The results indicated that the development of follicles in cultured chicken ovarian cortical pieces progressed smoothly and slowly similar to those in vivo, and the environment of chicken embryo was more suitable for the synchronous development of oocyte and follicular cells in the larger follicles. Meanwhile, the synchronous development could be more easily achieved in the chicken ovarian tissues of 12- to 15-wk chicken. Neither oocyte nor granulosa cells but the thecal cells of follicles developed in grafts of 13- to 15-d chicken within 10 d of culturing in ovo, and the CAM can ease follicular atresia in chicken ovarian grafts of 12- to 15-wk chicken. The proliferative performance of follicles in the grafts was not influenced by the environment of chicken embryo. These results indicated that the chicken embryo has the ability to support slow development of primordial and primary follicles in grafted ovarian cortical tissues of chickens of different ages. The CAM system may also prove useful for the study of early follicle development with further information.

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Isolation and culture of chicken primordial follicles

Leghari

Zhao

et al. 2015

Poultry Science

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The establishment of a primordial follicle culture system is important for the study of follicular development. Hence, the objective of this study was to isolate chicken primordial follicles and establish culture methods. Ovaries from 2-wk-old chickens were treated with trypsin-EDTA, collagenase II, or collagenase type IA, along with a mechanical isolation technique. Isolated follicles were cultured under different conditions. Results showed a significant difference in the follicular recovery and survival rates among different enzymes and methods used. The maximal follicular yield was obtained by trypsin+EDTA and collagenase II digestion, followed by collagenase type IA digestion. However, the highest follicular viability rate was observed in groups of collagenase type IA digestion and the mechanical isolation method. Enzymatic treatment resulted in higher misshapen oocytes or follicles, though the diameters of the follicles were not significantly changed. In addition, our follicle culture results for different conditions showed maximal survival rates of primordial follicles in alginate hydrogel beads after 12 d of culture. Thus, we successfully established methods for isolating and culturing chicken primordial follicles. The present method will greatly facilitate investigation of the regulation of follicular development.

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Effects of isolation and culture of turkey primary follicular oocytes on morphology and germinal vesicle integrity

Cited by 3 publications

References 12 publications

An Efficient Isolation Method for Domestic Hen (Gallus domesticus) Ovarian Primary Follicles

An Efficient Isolation Method for Domestic Hen (Gallus domesticus) Ovarian Primary Follicles

Development-promoting effect of chicken embryo membrane on chicken ovarian cortical pieces of different age

Isolation and culture of chicken primordial follicles

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