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Virginia Tech

1. Bolling, Laura Clayton. The Effect of Growth Hormone on Pig Embryo Development in Vitro and an Evaluation of Sperm-Mediated Gene Transfer in the Pig.

Degree: MS, Veterinary Medical Sciences, 2001, Virginia Tech

The objective of part one of this study was to determine if the presence of porcine growth hormone (pGH) during oocycte in vitro maturation (IVM) affected subsequent embryo development. Pig cumulus-oocyte complexes (COC) (n=987) were aspirated from slaughterhouse derived ovaries and cultured in BSA-free NCSU 23 medium containing porcine follicular fluid (10% v/v), cysteine (0.1 mg/ml) and hormonal supplements (eCG and hCG, 10 IU/ml each), 10 ng/ml EGF, and with or without pGH (100 ng/ml) for 22 h. The COC were then cultured in the same medium with or without 100 ng/ml pGH, but without hormonal supplements for an additional 22 h. After the completion of maturation culture, cumulus cells were removed and oocytes were co-incubated with frozen-thawed spermatozoa for 8 h. Putative embryos were transferred to NCSU 23 containing 0.4% BSA and cultured for 144 h. Embryo development was assessed on d 6 of culture. The treatment groups were as follows: treatment 1 = control group cultured in IVM medium alone; treatment 2 = 100 ng/ml pGH present of the first 22 h of maturation culture and absent for the second 22 h of maturation culture; treatment 3 = 100 ng/ml pGH absent for the first 22 h of maturation culture, but present for the second 22 h of maturation culture; and treatment 4 = 100 ng/ml pGH present throughout the entire IVM period. Embryos were visually scored for developmental stage at 144 h following fertilization. Each oocyte in the study received a developmental score, based on a scale of 1 = uncleaved, 2 = 2-cell embryo, 3 = 4- to 8-cell embryo, 4 = 9- to 16-cell embryo, 5 = morula, and 6 = blastocyst. The addition of pGH did not affect porcine embryo development as compared to the control (1.57 ± .08, 1.67 ± .08, 1.47 ± .08, and 1.60 ± .08, respectively; P > .10). Replicates within the study differed significantly from each other (P < .01) primarily because the development in replicate 6 was greater than for all others. There was a significant treatment by replicate interaction (P < .05); pGH added during the first 22 h of IVM and pGH added during the second 22 h of IVM in replicate 6 resulted in higher development scores than for controls and continuous pGH addition. However, in replicate 2, continuous pGH resulted in the greatest development. These results suggest that pGH may exert a stimulatory effect on embryo development when present in the IVM media; however, further studies using pGH in IVM culture are necessary. The objectives of the second part of the study were to examine aspects of intracytoplasmic sperm injection (ICSI) using membrane-disrupted spermatozoa, in vitro fertilization (IVF), and sperm-mediated gene transfer in the pig. Porcine oocytes were shipped overnight in maturation media at 39°C in a portable incubator. After 22 h of maturation culture, oocytes were washed in maturation medium without gonadotropins and cultured for an additional 22 h. Cumulus cells were removed and oocytes were divided into four treatment groups: treatment 1 = ICSI using membrane-damaged… Advisors/Committee Members: Pleasant, Robert Scott (committeechair), Velander, William H. (committee member), Knight, James W. (committee member), Gwazdauskas, Francis C. (committee member).

Subjects/Keywords: Embryo development; In vitro fertilization; Intracytoplasmic sperm injection; Transgenesis; Pig oocytes; Growth hormone; In vitro maturation

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6th Edition):

Bolling, L. C. (2001). The Effect of Growth Hormone on Pig Embryo Development in Vitro and an Evaluation of Sperm-Mediated Gene Transfer in the Pig. (Masters Thesis). Virginia Tech. Retrieved from http://hdl.handle.net/10919/35823

Chicago Manual of Style (16th Edition):

Bolling, Laura Clayton. “The Effect of Growth Hormone on Pig Embryo Development in Vitro and an Evaluation of Sperm-Mediated Gene Transfer in the Pig.” 2001. Masters Thesis, Virginia Tech. Accessed September 28, 2020. http://hdl.handle.net/10919/35823.

MLA Handbook (7th Edition):

Bolling, Laura Clayton. “The Effect of Growth Hormone on Pig Embryo Development in Vitro and an Evaluation of Sperm-Mediated Gene Transfer in the Pig.” 2001. Web. 28 Sep 2020.

Vancouver:

Bolling LC. The Effect of Growth Hormone on Pig Embryo Development in Vitro and an Evaluation of Sperm-Mediated Gene Transfer in the Pig. [Internet] [Masters thesis]. Virginia Tech; 2001. [cited 2020 Sep 28]. Available from: http://hdl.handle.net/10919/35823.

Council of Science Editors:

Bolling LC. The Effect of Growth Hormone on Pig Embryo Development in Vitro and an Evaluation of Sperm-Mediated Gene Transfer in the Pig. [Masters Thesis]. Virginia Tech; 2001. Available from: http://hdl.handle.net/10919/35823

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