Modified isolation and culture methods of human ovarian granulosa cells / 中国组织工程研究

BACKGROUND:To build up an effective method of isolating and culturing granule cels is a pivotal step to enhance fertilization-embryo transfer rate. Current studies mainly focus on the isolation methods of human ovarian granulosa cels rather than cel counting, purity and subsequent growth. OBJECTIVE: To establish the effective methods of isolating, purifying and culturing human ovarian granulosa cels in vitro. METHODS: Folicular fluid was harvested from women undergoing fertilization-embryo transfer procedures. Human ovarian granulosa cels were obtained from the folicular fluid by lysis treatment, precipitation method or density gradient centrifugation. Granulosa cel mucus masses were digested with type I colagen enzyme or hyaluronidase and then cultured in the culture medium with or without autologous folicular fluid. RESULTS AND CONCLUSION: Lysis treatment yielded the largest amount of granulosa cels compared to the precipitation method and density gradient centrifugation (P > 0.05,P < 0.05, respectively). Cels prepared by the three methods showed the same cel viability. After 24 hours of culture, the precipitation method obtained the largest amount of adherent granulosa cels (P < 0.05); and the density gradient centrifugation obtained the least amount of cels (P < 0.05). Compared with type I colagen enzyme, hyaluronidase took less time to digest the cels thoroughly. Autologous folicular fluid could promote the growth and survival of granulosa cels. These findings indicate that the precipitation method, though time-consuming, can obtain the highest cel viability and harvested the largest amount of granulosa cels after culture; hyaluronidase is more suitable for digesting granulosa cel mucus mass than type I colagen enzyme; autologous folicular fluid added into the culture medium is more conducive to granulosa cel growth.

A CLINICAL OBSERVATION OF THE EFFECT OF "MUYINGLE" ON LACTOGENESIS / 中国海洋药物

Thirty mothers of four dadys later after delivery with agalactia, whose babies needed additional milk over one third of physiological needs, or agalorrhea were chosen to take 125 g of MUYINGLE divided into 3 parts a day for observing its lactigenous effect, and thirty other agalactous mothers, who might chose any kind of traditional Chinese lactagogue foods to eat, as control group. Those subjects whose babie's additional milk was less than one fourth or half of physiolgical needs for agalactia or agalorrhea respectively after four days with MUYINGLE were effectual. The results showed that the lactagogue efficacious rate of MUYINGLE and control group were 86. 7% and 33. 3%, respectively. The lactagogue effects were significant difference between the two groups (P