• Korean Journal of Animal Reproduction
• /
• v.25 no.2
• /
• pp.163-169
• /
• 2001

OSBA(oocytes-sperm binding assay) is a tool developed for rapid test of optimal condition of IVF medium and protein source by binding ability of mouse sperm and egg. Mouse oocyte-cumulus complexes were prepared by removing of the cumulus cells with 0.1% hyaluronidase. 10$\pm$2 oocytes per 30 ${mu}ell$ medium drop were inseminated with 3 ${mu}ell$ sperm suspension and were cultured f3r 3 hours and 24 hours, respectively. And the oocytes were recovered gently and the No. of sperm bound on oocytes were counted. In the Exp. 1, the ratio of oocytes bound with one sperm at least were 60.2%(50/83), 2%(2/77) and 100%(79/79) in the medium with no protein, FBS(15%, v/v) and BSA(0.4%. w/v), respectively, Fetal bovine serum(FBS) seriously inhibited sperm binding on oocyte, although bovine serum albumin(BSA) promoted the binding ability. The inhibiting effect of FBS was dependent on the concentration of FBS. The sperm binding ability according to oocyte maturity was tested in the Exp. 2. There was no significant difference between Met. II (mature) and Met. I (intermediate mature) oocytes in the number of oocytes bound with sperm and the number of sperm bound on oocytes. Finally, in Exp. 3, two batches of Ham's F10 medium with good and poor quality by OSBA were tested (The ratios of embryos developed from PN 1-cell stage to hatched blastocyst; 25% vs. 70%). In the medium with good quality, sperm binding ability was significantly increased (P ＜ 0.05). The ratio of oocytes bound with one sperm at least was 66% and 90% in the medium with poor and good quality, respectively. Conclusively, It was possible to test IVF medium condition rapidly and easily by OSBA.

• Korean Journal of Medicinal Crop Science
• /
• v.2 no.1
• /
• pp.60-66
• /
• 1994

This study was conducted to determine the optimum conditions of inducing callus, proliferating callus, forming somatic embryos, and regenerating plantlets via somatic embryogenesis, for the purpose of producing artificial seeds and substantially developing plant factory technologies that can be employed to all seasons production of Bupleurum plants. Callus was efficiently induced from leaf tissues at three leaf stage in the MS medium supplemented with 2, 4-D 2mg /1 and thidiazuron(TDZ) 0.lmg /1. Callus induction from leaf tissues at maturity was mostly effective in the mixture of 2,4- D 2mg /1 and TDZ 1.0mg /1 while that from flower bud tissues was fairly good in the MS medium containing 2,4-D 1 or 2mg /1.Callus was formed in 15 to 20 days after culture initiation in the MS media supplemented with 2, 4- D 1-2mg /1 and TDZ 0.l-1.0mg /1. Such hormones as kinetin 3mg /1, GA 1mg /1, and the mixture of GA 1mg /1 and TDZ 1mg /1 effected markedly to proliferate the callus cells.The optimum temperature and light intensity for callus culture were found to be $25^{\circ}C$ and 3000 Lux, respectively. Direct plant regeneration from cultured callus was fairly made on hormone-free MS or half-strength MS medium. Somatic embryogenesis was most frequently observed in hormone-free media:60 somatic embryos per 20ml in MS medium and 28 somatic embryos per 20ml in half -strength MS medium. There were three stages-globular, heart, and torpedo-in development of somatic embryos, among which globular stage was more frequently observed in MS medium rather than in half-strength MS medium. Somatic embryos induced from suspension culture fairly differentiated a number of shoots and roots on hormone-free and half-strength MS solid medium.