This research was conducted to obtain the basic information on the cell block phenomenon occuring during early development in vitro of mouse embryos. Early embryos were recovered at 3h post-hGG injection(hph). Various chemicals (EDTA, EGTA, DTPA, MA and PRA) were tested to examine the effects of them on the overcoming the 2-cell block phenomenon. One hundreds M of the chelating agents were added to the M16 medium containing embryos. The treated embryos were worked and transferd to fresh M16 medium after 1, 3, 6 and 12h of treatment. Development was examined at 58 and l2Oph injection, respectively. 44.7~68.9% of the treated embryos developed to 4-cell stages at 58hph. Only 17.6~60.3% of the embyos developed upto blastocyst at l20hph. Whereas control embryos showed slightly lower development in M16 medium alone (38.9~42.4%, 4-cell and 3.8~65.5%, blastocyst). Three mitogenic agents were tested. 51.6~63.8% and 43.4~48.1% of embryos developed up to 2-cell and blastocyst stage, respectively when treated in 5 g PHA-M Imi for 5 min, 1, 3 and 6h subsequently cultured in fresh M16 medium. Control embryos only showed 38.8% for 4cell and 5.9% fo blastocyst at 58 and l2Ohph, respectively. 100M PMA was also beneficial for the 2-cell block. Showing better development them that of control (42.4 vs 57.9~59.4% 4cell and 5.9 vs 25.0~55.6% blastocyst, respectively. However 1M butyric acid was toxic to early embryos, thus arresting further development. These result indicate that either chelating or mitogenic agents could be used to overcome the "in vitro 2-cell block" occuring during early development in vitro of ICR embryosCR embryos

This experiment was investigated the effect of presence of granulosa cells from follicles of different size on bovine oocyte maturation, cleavage and development to late stage. The nuclear and cytoplasmic maturation of oocytes in the IVM-IVF system are critical for subsequent embryo development. Granulosa cells when the co-cultured with oocytes may interact with cumulus-oocytes complexes and influence the development competence of the oocytes. Granulosa cells from medium (2~6 mm) and large(>1O mm) size follicles were recovered by aspiration, washed 3 times by centrifugation at 500 x g for 5 min. and used for co-culture at a concentration of 2~3 x 106 cells/mi. The oocytes were matured in vitro (IVM) for 24 hrs. in TCM-199 supplemented with 35 g/ml FSH, 10 g/ml LH, 1 g/ml estradiol-17 and granulosa cells at 39 under 5% in air. They were fertilized in vitro (IVF) by epididymal spermatozoa treated with heparin for 24 hrs., and then the zygotes were co-cultured in vitro (I VC) with bovine oviductal epithelial cells for 7 to 9 days. The assessment of maturation revealed that Grade J oocytes showed significantly(P

본(本) 연구(硏究)는 들깨 국내 육성종을 포함한 수집종(蒐集種)중 20종(種)을 공시하여 이들 간의 형태적(形態的)인 주요(主要) 특성(特性), 생화학적(生化學的)인 특성(特性)과 세포유전학적(細胞遺傳學的)인 특성(特性)을 조사(調査)하고 유전적(遺傳的) 다양성(多樣性)을 밝혀 들깨의 분류(分類)와 품종육성(品種有成)에 있어서의 중요(重要)한 기초자료(基礎資科)를 얻고자 본(本) 실험(實驗)을 수행(遂行)하여 얻은 결과(結果)를 요약(要約)하면 다음과 같다. 들깨 수집종의 형태적(形態的), 생화학적(生化學的), 세포유전학적(細胞遺傳學的) 특성(特性)을 조사(調査)해 본 바, 형태적(形態的) 특성(特性)으로 종피색은 약간의 농도차이는 있었으나 회갈색(灰褐色), 갈색(褐色), 암갈색(暗褐色)(흑색)등 3군(群)으로 나눌 수 있었으며, 우립중(于粒重)은 3.5g 이하인 것이 11종(種)으로 가장 많았고 4.5g 이하인 것이 4종(種) 4.5g 이상인 것이 5종(種)이었다. 또한 줄기색은 적색(赤色)과 녹색(綠色)으로 분류되었는데 적색(赤色)은 안동재래를 포함 5종(種)이었고 나머지는 녹색(綠色)이었다. 생화학적(生化學的)인 특성(特性)으로 각 수집종(蒐集種)을 대상으로 지방산(脂肪酸) 조성을 조사(調査)해 본 바, 불포화도가 가장 높은 linolenic acid는 53%(영양재래)에서 65%(의성재래)에 이르기 까지 다양(多樣)하였으며, 59-61% 범위에 속하는 종(種)이 12종(種)으로 가장 많았다. 염색체(染色體) 수(數)는 조사한 4개의 수집종(蒐集種)(안동재래, 엽실, 영광재래, 대영) 모두가 2n=38로 나타났다.

The studies on the carried out to investigate the effects of co-culture with uterine fluids and uterine epithelial cells on the in-vitro fertilization and developmental rate of porcine follicular oocytes. The ovaries were obtained from slaughtered swine. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluids from the visible follicles of diameter 3~5 mm. The follicular oocytes were cultured in TCM-199 medium containing hormones and 10% ECS for 46~48 hrs in a incubator with 5% in air at 38.5 and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation of heparin. The results obtained in these experiments were summarized as follows ; 1.The in-vitro maturation and fertilization rate of porcine oocytes co-cultured with uterine fluids in TCM-199 medium were 68.0% arid 55.7%, the rates were higher than of control, 56.5% arid 38.7%. 2. When the in-vitro fertilized oocytes were co-cultured with porcine uterine epithelial cells, the fertilization rate was 60.3%, the rates were higher than that of control, 35.7%. 3. When the in-vitro fertilized oocytes were co-cultured with porcine uterine epithelial cells, the development rate to be blastocyst was 12.4%, the rates were higher than that of control, 9.2%(p<0.05).

To investigate the effect of extracellular matrix proteins on the in vitro development of ethanol-induced parthenogenetic eggs of ICR strain mice, those were cultured in vitro in fibronectin, gelatin, or collagen precoated culture dishes containing 1.5 ml of NaH-C03-BMOC-3 medium at 37 for 96 hrs. under the atmosphere of 5% and 95% air. Fibronectin, gelatin, or collagen significantly(P1.4, 45.4i1.4, and 44.8O.9, respectively. And the diameter of those eggs ranged 104.61.9, 102.82.3, and 103.4O.8 m, respectively.