Although the efforts to establish fish embryonic stem cells (ESCs) have been made for a long time, derivation of authentic ESCs that possess pluripotency is still difficult suggesting a need for the stepwise optimization of the methods to establish fish ESCs. Primary culture of the blastomeres from the embryos at blastula stage is a critical step for establishing continuous ESC lines. Here, we evaluated the effects of temperatures and basal media on primary culture of blastula embryo-derived blastomeres in marine medaka (Oryzias dancena). The blastomeres were isolated from the blastula embryos and cultured in various conditions designed by the combination of 4 temperatures including 28°C, 31°C, 34°C, and 37°C and 2 basal media including Dulbecco’s modified eagle’s medium (DMEM) and Leibovitz’s L-15 medium (L15). With the exception of a case cultured in L15 at 31°C, the rate of primary cell adherence reached 100% when the blastomeres were cultured over 31°C. The period for primary adherence was significantly shorter in the groups cultured in 34°C and 37°C than in the ones in 28°C and 31°C. The proportion of subculture was significantly high in the group cultured in DMEM at 31°C compared to the other groups. Collectively, we demonstrated that the culture in DMEM at 31°C was effective to primary culture of the blastomeres derived from blastula embryos.

Dendrobium kingianum의 유묘증식(幼苗增殖)을 위한 기초자료(基礎資料)를 얻고자 발아(發芽)에 미치는 기본(基本) 배지(培地)의 영향(影響)과 유묘생장(幼苗生長)에 미치는 hyponex의 종류별농도(種類別濃度), auxin 농도(濃度), 당(糖), 한천(寒天) 및 pH의 영향(影響)을 알고자 실험(實驗)한 결과(結果)를 요약(要約)하면 다음과 같다. 종자(種子)의 발아(發芽)는 hyponex +peptone 가 함유된 배지(培地)에서 가장 양호(良好)하였으며 유묘(幼苗)의 증식(增殖) 및 생장(生長)은 hyponex +peptone 가 함유된 배지(培地)에 IBA , sucrose , agar 첨가배지(添加培地)에서 multiple shoot가 형성(形成)되었으며 유묘(幼苗)의 생장(生長)을 촉진(促進)시키기 위해서는 auxin이 첨가(添加)되지 않은 배지(培地)에 sucrose 한천(寒天) 및 pH는 5.0이 가장 적합(適合)하였다.

양하(襄荷)를 대양번식(大量繁植) 하고자 경정부위(莖頂部位)의 기내배양(器內培養) 방법(方法)을 확립(確立)하기위하여, 경정부위(莖頂部位)의 생장(生長)에 알맞은 기본부지(基本部地) 및 식물생장조절제(植物生長調節劑)의 농도(濃度)를 구명(究明)하고자 본(本) 실험(實驗)을 수행(遂行)하였던 바 그 결과(結果)를 요약(要約)하면 다음과 같다. 1. 양하(襄荷)의 경정부위(莖頂部位)는 치상후(置床後) 10~12주(週) 후(後)에 기관(器官) 분화현상(分化現像)이 나타나서 shoot와 root가 발생(發生)되었다 2. 양하(襄荷)의 경정(莖頂)은 MS기본부지(基本部地)에 치상(置床)한 환경(環境) 비교적(比較的) 생장(生長)이 잘 되었다. 3. 기내(器內)의 Plantlet의 생장(生長)은 식물생장조절제(植物生長調節劑) NAA 0.5ppm과 BA 5.0ppm의 혼합(混合) 첨가배지(添加培地)에서 좋은 경향(傾向)이었다.

The study was conducted to determine the optimal basal media and the concentration of plant growth regulator for germination of seeds and growth of plantlet from Dendrobium monile. The results obtained were summarized as follows. Germination was similar in light and dark condition, but the growth of plantlet after germination was better under dark than under light condition in several media. Germination was best in Hyponex and Kyoto solution medium among the 9 media tested. The number of roots/shoot was most in the Hyponex medium containing 0,1ppm NAA and 1.Oppm BA.