| The ovarian follicle is the basic structural and functional unit of mammalian ovary and it provides the microenvironment for the growth and maturation of oocyte. Despite the large population of primordial follicles present in the ovary during folliculogenesis, many follicles undergo atresia, and only a few of them develop to the preovulatory stage, which is just a waste of hereditary resources. The isolation, culture and the in vitro maturation (JVM) of porcine preantral follicles were studied in order to empolder and utilize these resources. The objective of this study is to empolder further porcine genetic potential and provide large number of oocytes in high quality for embryo biotechnology such as in vitro fertilization, cloning and transgenic technology.This study compared the effects of two mechanical methods on the isolated porcine preantral follicles regarding the number, handling times of isolation preantral follicles and their livability after three days culture in vitro as indexes. The result showed that needle scraping method harvested significantly more porcine preantral follicles(Φ < 50um: 203600±59000, 50μm□Φ□150μm: 29.40±10.34, Φ>150μm: 9.30±3.29) than cutting(Φ<50um: 38900±12000, 50μm□Φ□50μm: 13.95±3.70, Φ150μm: 4.95±1.61) ( p<0.01). The average handling time of needle scraping method for each ovary (16.16±1.34min) was less than that of cutting (20.30±1.48min) ( p<0.01). The livability of preantral follicles after three days in-vitro culture was not significant different between the two groups (81.00% vs 78.97%, p>0.05). The results indicated needle scraping mechanical isolation could be used to maintain normal physiological functions of preantral follicles as it kept the natural figures of oocytes and granulosa cells and preserved the integrity of basement membrane.Two basic medium of Ml99 and NCSU23 were compared as their effects on the grow speed, antrum formation and follicular livability. These results demonstrated that preantral follicles cultured in M199 medium grew faster than those mNCSU23 (5d: 31.01±15.79 vs21.73±12.69, p<0.01,10d: 44.69±25.90 vs 33.25 ±17.35, p<0.05) . The follicular growth rate during the first 5 days of culture was higher than that during the latter 5 days in both of the two culture-medium groups (M199: 31.01±15.79 vs 13.68±11.50, p>0.01;NCSU23:21.73±12.69 vs 11.52±9.19, p< 0.01) . However, antrum formation and follicular livability had no difference between the two groups. Therefore, M199 was chosen to investigate the effects of different culture methods and various concentrations of FSH on the in vitro culture (IVC) of porcine preantral follicles.96-well plates, hollowness and granulose cells coating culture systems were adopted in culturing porcine preantral follicles in vitro. The results showed that the follicular growth rate in hollowness culture system was lower than that in 96-well plates(5d: 26.44±17.38 vs 19.18±9.37, p...
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