Study On In Vitro Development Of Oocytes And Morphology Of Viable Ovarian Follicles Of Human

The morphology and developmental regulating factors on human ovarian follicles were studied in this paper, involving follow aspects: 1. Morphological characteristics of human ovarian foilicles With the method of ovarian tissue digested by collagenase, the distribution of primordial follicles, primary follicles and secondary follicles in cortex, the relation between ovarian follicular number and ages, the variation of ovarian follicular growth and atretic were investigated, compared and analyzed. The diameters of primordial follicles and primary follicles ranged 30挆64 Ii m and 60?9 it m respectively. The primordial follicles with a diameter <35 ji m were only 2.32% of all. The diameter of more than 76% of primordial follicles and more than 80% of primary follicles were 40 -~ 54 it m and 65 憲 84 ji m respectively. The follicular number decreased with the age increasing. The follicular numbers were 63l.43?25.39~. 303.17?54.61 and 70.38?7.99 in 1/4 of ovary for the 20---year~ 30---year and 40----year groups respectively. There were significant differences among the three groups (P<0.01). After 50 years, the number of follicles decreased significantly(P<0.001) , only was 13.5 + 9.67. 80 h~il~: A 1*~F~W ~ The follicles distributed different in cortex with different ages. The follicles distributed in cortex with even density in 20 years group. Non-follicle-area appeared and remaining follicles distributed in cluster in 30~梱ears group. The non-follicle-area increased to form larger area and only a few follicles scattered in cortex in 40拁-years group. There are very few follicles in postmenopausal woman group. The primordial, primary and secondary follicles located in the outer, middle and inner cortex respectively. The early antral follicles located in more inner cortex. The secondary follicles and early antral follicles grew by blood vessels and in cluster. 2. Study on isolation of human ovarian follicle Four isolation methods, mechanical pipetting, collagenase digestion, microdissection and the combination of collagenase digestion and microdissection were compared. Isolation times were 44.56 ?6.26mm, 48.25?.33 mm and 185.38?5.91mm and obtained follicles were 5.19?.16, 4.44?.59 and 33.25 ?12.89 for mechanical pipetting, collagenase digestion, microdissection and the combination of collagenase digestion respectively when isolating the 1/4 of ovary. The isolated follicles most were primordial follicles in mechanical pipetting and collagenase digestion groups. More follicles and three stage follicles including primordial, primary and secondary follicles were isolated in microdissection and the combination of collagenase group in spite of its time consuming (P < 0.01 ) . The isolated follicles grew and secreted E2 normally. 3. Study on in vitro culture of human follicle When follicles cultured in control group and FSH groups with concentration of 0.5 IU/mL, 1.0 IU/mL ~~1~lJ 2.0 IU/mL, the survival times of primordial follicle were 4.41 ?l.96d, 4.31 ?1.66d 454+ l.96d and 4.25?l.83d. The survival times of primary follicle were 4.69?.02d, 4.13?.96d, 4.36?.17d and 4.47 ?.07d. The survival times of secondary follicle were 2.80?l.69d, 5.36?.63d, 5.47+2.50d and 8.13 ?.l9d. There was a positive correlation between survival time and concentration of FSH in secondary follicle gr...