title: The effects of human recombinant granulocyte-colony stimulating factor treatment during in vitro maturation of porcine oocyte on subsequent embryonic development.
abstract: Granulocyte colony-stimulating factor (G-CSF) is required for proliferation, differentiation, and survival of cells. It is also a biomarker of human oocyte developmental competence for embryo implantation. In humans, the G-CSF concentration peaks during the ovulatory phase of the ovarian cycle. In this study, the expressions of G-CSF and its receptor were analyzed by polymerase chain reaction in granulosa cells (GCs), CL, cumulus cells (CCs), and oocytes. Cumulus-oocyte complexes were aspirated from antral follicles of 1 to 3 mm (small follicles) and 4 to 6 mm (medium follicles). Cumulus-oocyte complexes from two kinds of follicles were matured in protein-free maturation medium supplemented with various concentrations of G-CSF (0, 10, and 100 ng/mL). By real-time polymerase chain reaction, the expressions of G-CSF and its receptor were detected in GCs, CL, CCs, and oocytes. Interestingly, the G-CSF transcript levels were significantly lower in oocytes than in the other cell types, whereas the G-CSF receptor transcript levels in oocytes were similar to those in GCs. After 44 hours of IVM, no differences in the rate of nuclear maturation were detected; however, the intracellular reactive oxygen species levels in oocytes from both groups of follicles matured with 10 ng/mL of human recombinant G-CSF (hrG-CSF) groups were significantly lower (P < 0.05). After parthenogenetic activation, the cleavage rates were significantly (P < 0.05) higher in 100 ng/mL hrG-CSF-treated small (63.3%) follicles than in 0, 10 ng/mL hrG-CSF-treated small (38.6% and 49.0%, respectively) follicles and 0 ng/mL hrG-CSF-treated medium (52.1%) follicles, and the cleavage rates were significantly (P < 0.05) higher in 10 ng/mL hrG-CSF-treated medium (76.3%) follicles than in all other groups. The blastocyst formation rates were significantly (P < 0.05) higher in 100 ng/mL hrG-CSF-treated small (31.2%) follicles than in 0 and 10 ng/mL hrG-CSF small (10.4% and 15.6%, respectively) follicles, and the 10 ng/mL hrG-CSF medium (45.7%) follicle was significantly (P < 0.05) higher than in all other groups. The total cell number in blastocysts from the 10 ng/mL hrG-CSF medium (106.5) follicles was significantly (P < 0.05) increased compared to 0, 10, 100 ng/mL hrG-CSF small (55.0, 73.7 and 59.5, respectively) follicles and 0, 100 ng/mL hrG-CSF-treated medium (82.5 and 93.5, respectively) follicles. After IVF, the blastocysts stage was significantly (P < 0.05) increased in 10 ng/mL hrG-CSF-treated medium (36.4%) follicles. Fertilization efficiency was significantly high in 100 ng/mL of small (29.1%) and 10 ng/mL of medium (44.0%) follicles. We also examined the Bcl2 and ERK2 transcript levels and found that they were significantly higher in the small and medium follicle treatment groups. In conclusion, these results indicate that hrG-CSF improve the viability of porcine embryos.
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, South Korea.
authors.forename: Lian
authors.initials: L
authors.lastname: Cai
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, South Korea.
authors.forename: Yubyeol
authors.initials: Y
authors.lastname: Jeon
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, South Korea.
authors.forename: Junchul David
authors.initials: JD
authors.lastname: Yoon
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, South Korea.
authors.forename: Seon-Ung
authors.initials: SU
authors.lastname: Hwang
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, South Korea.
authors.forename: Eunhye
authors.initials: E
authors.lastname: Kim
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, South Korea.
authors.forename: Kyu-Mi
authors.initials: KM
authors.lastname: Park
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, South Korea.
authors.forename: Kyu-Jun
authors.initials: KJ
authors.lastname: Kim
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, South Korea.
authors.forename: Ming Hui
authors.initials: MH
authors.lastname: Jin
authors.affiliation: Department of Theriogenology, College of Veterinary Medicine, Kangwon National University, Kangwon, South Korea.
authors.forename: Eunsong
authors.initials: E
authors.lastname: Lee
authors.affiliation: Department of Biotechnology, School of Life Sciences and Biotechnology, Korea University, Seoul, South Korea.
authors.forename: Hyunggee
authors.initials: H
authors.lastname: Kim
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, South Korea.
authors.forename: Eui-Bae
authors.initials: EB
authors.lastname: Jeung
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, South Korea. Electronic address: shhyun@cbu.ac.kr.
authors.forename: Sang Hwan
authors.initials: SH
authors.lastname: Hyun
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology; D054885:Cumulus Cells / Q000737:chemistry; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D015870:Gene Expression; D016179:Granulocyte Colony-Stimulating Factor / Q000032:analysis / Q000235:genetics / Q000494:pharmacology*; D006107:Granulosa Cells / Q000737:chemistry; D006801:Humans; D059471:In Vitro Oocyte Maturation Techniques / Q000662:veterinary*; D009865:Oocytes / Q000737:chemistry; D012333:RNA, Messenger / Q000032:analysis; D017382:Reactive Oxygen Species / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction; D016188:Receptors, Granulocyte Colony-Stimulating Factor / Q000032:analysis / Q000235:genetics; D011994:Recombinant Proteins; D013552:Swine
chemical_list: D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D016188:Receptors, Granulocyte Colony-Stimulating Factor; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor
keywords: Follicle size; Granulocyte colony-stimulating factor; IVM; Oocyte; Porcine