The present study examined changes in surface shape and pore size observed in carbon black particles isothermally oxidized in an air atmosphere according to their burn-off ratio. Carbon black materials were fed into a horizontal tubular furnace in an air atmosphere when the inside temperature reached 600 °C. Subsequently, while changing the isothermal oxidation time, carbon black samples with different burn-off ratios were obtained, i.e., 10.5, 20.0, 30.4, 41.0, 49.9, 59.8, 71.1, and 81.0%. The scanning electron microscope analysis revealed that the observed carbon black particles were in the form of aggregated primary particles, and that there was no change in the particle size of these primary particles as the burn-off process proceeded. The latter observation supported the observation that pores were formed in the carbon black samples during the burn-off process. Notably, the Brunauer–Emmett–Teller analysis exhibited hysteresis curves, indicating that the corresponding adsorption isotherms were of IV-type. It was also found that the area of the hysteresis curves increased as the burn-off process proceeded. The specific surface area of the raw carbon black sample was 58.00 m2/g, while that of the 81.0% sample was about 4.1 times the figure at 240.27 m2/g. The total pore volume VT was 0.17 cm3/g for the raw sample, and it was much higher for the 81.0% sample at 0.58 cm3/g. The transmission electron microscope analysis showed that the raw carbon black particles had a spherical shape with a smooth surface, but inner pores were not observed. In the 49.9% sample, pores with a size of about 5 nm were observed inside carbon black particles. Notably, the size of the pores observed in the 81.0% sample was about 20 nm and the large pores were created by the collapsing and merging of the smaller pores by oxidation.

Despite that porcine spermatogonial stem cells (pSSCs) have been regarded as a practical tool for preserving eternally genetic backgrounds derived from pigs with high performance in the economic traits or phenotypes of specific human diseases, there were no reports about precise definition of niche conditions promoting proliferation and maintenance of pSSCs. Accordingly, we tried to determine niche conditions supporting proliferation and maintenance of undifferentiated pSSCs for short-term. For these, undifferentiated pSSCs were progressively cultured in different composition of culture medium, seeding density of pSSCs, type of feeder cells and concentration of growth factors, and then total number of and alkaline phosphatase (AP) activity of pSSCs were investigated at post-6 day culture. As the results, the culture of 4x105 pSSCs on mitotically in activated 2x105 STO cells in the mouse embryonic stem cell culture medium (mESCCM) supplemented with 30 ng/ml glial cell line-derived neurotrophic factor (GDNF) was identified as the best niche condition supporting effectively the short-term maintenance of undifferentiated pSSCs. Moreover, the optimized short-term culture system will be a basis for developing long-term culture system of pSSCs in the following researches.

There are diverse methods of cryopreservation of mammalian embryos with variable degrees of success. Although cryopreservation technique of mammalian embryos has been advanced, freezing stress affect to cellular event such as apoptosis and autophage in embryos. The objective of the study is to investigate the affection of to survival, development, live offspring, apoptosis and autophagy on embryo. Mouse embryos were vitrified and thawed using normal straw and modified cut standard straw (M-CSS), then in vitro cultured until blastocyst stage and transferred to recipient. Recovery rates (100 vs 99.2%), survival rates (99.2 vs 78.6%), developmental rates (18.4 vs 10.7%), total cell numbers (45 vs 37), preganacy rates (34.5 vs 25%) and offspring numbers (10.1 vs 4.9 %) of M-CSS group are significantly higher than those of normal straw vitrified group. Also, rate of apoptosis in blastocysts developed using M-CSS (1.9%) was significantly lower than using normal straw vitrification (2.7%). Apoptosis-related gene, caspase 3, was expressed at the highest level in blastocysts derived from normal straw group. However, no differences of autophagy related gene, Atg6 and expression of LC3 between normal straw and M-CSS groups were observed. In conclusion, the standard vitrification procedure induces mitochondrial apoptosis in zygotes in an autophagy-independent manner, whereas the novel M-CSS procedure may improve embryo vitrification.