포도 ‘My heart’의 기내증식과 기외이식 후 생장에 있어서 배양용기에 부착한 미세공극 Filter 처리가 건전한 유묘를 생 산하는데 효과적인 방법을 찾고자 실시하였다. 미세공극 Filter는 환기구 크기별로 White filter type (50.0 mm×3.5 mm)과 Green filter type (50.0 mm×7.0 mm) 으로 구분하여 밀폐 처리와 비교하였다. Shoot tip 배양에 있어서 Green filter type에서 shoot 분화율이 75%로 White filter type 77% 와 밀폐처리 80% 보다 낮았지만 투명화 shoot 발생율은 4% 로 White filter type 13.4%, 밀폐처리 24.7%에 비하여 9.4－ 20.7%가 적었다. 전체 배양묘의 54.9%가 투명화 발생이 되 었을 때 Green filter type으로 90일 동안 계대 배양하고 조사 한 결과 투명화율은 11.8%로 감소하였고 분화한 shoot 수는 89개에서 915개로 증가하였다. Filter type에 따라 IBA 2.0 mg·L-1를 첨가하여 30일 동안 배양을 하였을 때 Green filter type에서 기내 발근율 100%, 뿌리 수 7.3개, 엽수 10.0개로 White filter type과 밀폐 처리보다 좋았다. 기외 이식하고 15 일 후의 유묘 생존율도 Green filter type에서 100%로 다른 처 리에 비하여 1.5－29.5% 더 높았고 초장이 11.0cm, 생체중이 1.7g 으로 가장 양호하였다. 미세공극 Green filter 처리는 포 도 ‘My heart’의 기내배양에서 shoot 투명화를 감소시키고 shoot와 뿌리 생성을 촉진시키고 기외이식 후 생장은 통계적 으로 유의하게 확인되어 건전한 유묘 생산에 효과적이었다.

An efficient method for in vitro propagation and growth of the wild garlic(Allium ochotense Prokh.) was established. Bulbs of wild garlic were collected from Ullung Island, Korea, and the growth pattern of plantlets on various culture media was observed. High growth of shoot was obtained on LP, NN and N6 medium. The growth media supplemented with 3%(w/v) sucrose was found optimal for shoot growth. After 10 weeks multiple shoots were observed in the plantlets growing on the medium containing 1.0 mg/l of zeatine and 0.1 mg/l NAA. Roots were induced directly at the base of the shoot in all treatments. The medium with 2.0 mg/l of IBA proved to be the best rooting medium. The studies of this kind may be used to develop strategies for large-scale propagation of elite wild garlic.

The PHBV nonofibrous membrane fabricated by electron-spinning method for tissue engineered bone regeneration scaffold was evaulated in terms of cellular prolieration and cryopreservation efficiency. The rat calvarial periosteum derived primary cells were cultured with PHBV nanofibrous membranes and analyzed the cellular proliferation and differention fashion and cryopreservation potential by in vitro MTT assay as well as ALP staining and Alizarine red staining with or without cryopreservation for 2 weeks. The rat calvarial periosteum derived primary cells cultured with PHBV nonofibrous membrane showed favorable proliferation and alkaline phosphatase activity with numerous mineral nodule formation regardless of cryopreservation, even though its efficiency was slightly decreased in cryopreserved condition. These findings suggest that PHBV nanofibrous membrane can be applicable as an efficient cell engineered membrane for guided bone regeneration or scaffold for tissue engineered bone regeneration.

Royal jelly (RJ) is exclusive food that is secreted from the hypopharyngeal and mandibular glands of worker honeybees, and it is well known to be a necessary for the growth of the queen honeybee Although fresh royal jelly have been demonstrated to enhance wound healing, the wound healing effects of water soluble royal jelly (WSRJ) have not been elucidated. We investigated whether WSRJ promotes the migration, attachment, and proliferation of human dermal fibroblasts (HDFs) during in vitro wound healing. HDFs were treated with 1-5ug/ml WSRJ and RJ for up to 24hr following wound formation. Cell migration was assessed by measuring recovery from wound margin, while cell attachment and proliferation were determined by MTT assay. By observing the numbers of cell attached, we confirmed that not only WSRJ but also RJ did not affect on the initial cell adhesion. WSRJ (5 ug/ml) enhanced cell migration rate approximately 84.3% in HDFs at 24hr, whereas RJ (5 ug/ml) increased cell migration rate 71.3% in HDFs at 24hr, which is similar to cell migration rate of WSRJ 1 ug/ml (73.7%). In cell proliferation assays, WSRJ induced an increase in the number of HDFs, compared with control and RJ. In conclusion, WSRJ promotes cell migration with increased cell proliferation in an in vitro wound healing model.

Human fibroblasts that maintain the structural integrity of connective tissues by secreting precursors of the extracellular matrix are typically cultured with serum. However, there are potential disadvantages of the use of serum including unnatural interactions between the cells and the potential for exposure to animal pathogens. To prevent the possible influence of serum on fibroblast cultures, we devised a serum-free growth method and present in vitro data that demonstrate its suitability for growing porcine fetal fibroblasts. These cells were grown under four different culture conditions: no serum (negative control), 10% fetal bovine serum (FBS, positive control), 10% knockout serum replacement (KSR) and 20% KSR in the medium. The proliferation rates and viabilities of the cells were investigated by counting the number of cells and trypan blue staining, respectively. The 10% FBS group showed the largest increase in the total number of cells (1.09 × 105 eell₃/ml). In terms of the rate of viable cells, the results from the KSR supplementation groups (20% KSR:64.7%; 10% KSR: 80.6%) were similar to those from the 10% FBS group (68.5%). Moreover, supplementation with either 10% (30 × 104 eell₃/ml) or 20% KSR (4.8 × 104 cells/ml) produced similar cell growth rates. In conclusion, although KSR supplementation produces a lower cell proliferation rate than FBS, this growth condition is more effective for obtaining an appropriate number of viable porcine fetal fibroblasts in culture. Using KSR in fibroblast culture medium is thus a viable alternative to FBS.

1. 생쥐 고환으로부터 얻은 세포를 배양하여 군집을 형성하는 것을 관찰할 수 있었으며, AP, SSEA-1, -3, -4과 Integrin 6, 1 및 Oct4의 발현을 확인하였다. 2. 생쥐 생식줄기세포를 3-5일정도 배양하게 되면, 여러 층으로 이루어진 군집을 이루게 되는데 이는 생쥐 배아줄기세포나 배아생식줄기세포의 형태와 같은 것이었다. 3. 생쥐 생식줄기세포를 체외에서 효과적으로 분리, 배양할 수 있는 조건을 확립하였다.

This study was performed to develop the mass propagation system using tissue culture technique to supply the seeds of Elephant garlic (Allium ampeloprasum L.) which has difficulty in propagation. Immature spathe of Elephant garlic was cultured on Murashige & Skoog (MS) medium supplemented with two plant growth regulators, naphthaleneacetic acid (NAA) and kinetin. After 6 weeks of culture, the highest number of shoot (14.9/explant) was obtained when the immature spathe with 10 ㎝ length was cultured right after harvesting. In MS medium supplemented with 2 ㎎/L kinetin and 0.5 ㎎/L NAA, the most vigorous growth characteristics was observed, the shoot number was 14.9/explant, its length was 11.3 ㎝, and its fresh weight was 2.5 g. When the bulblets were cultured in MS medium with 2 ㎎/L kinetin and 0.5 ㎎/L NAA, the addition of 30 ㎎/L adenine improved their proliferation and growth significantly, the highest bulblet formation rate (48%) was obtained. The addition of 7% sucrose also increased the bulblet formation rate at the highest frequency of 98.2%. The shoots were shown be more vigorously proliferated at the secondary subculture stage rather than primary culture stage, their propagation rate was 80% after subculture.

본 실험은 여우구슬의 기내 부정근 유도 및 증식조건의 확립을 목적으로 수행되었다. 우선 여우구슬의 기내 발아체로부터 부위를 달리하여 부정근을 유도한 결과 줄기부위는 뿌리보다 양호한 부정근의 유도를 보였다. 또한 유도된 부정근을 이용하여 옥신의 종류(IAA, IBA, NAA와 2.4-D)에 따른 부정근 유도율을 조사한 결과 IBA와 NAA는 IAA와 2.4-D보다 높은 유도율을 보였다. IBA의 농도에 따른 유도율과 증식효율은 IBA가 0.5 mg/L첨가되었을 때 가장 높은 유도 및 증식효율을 보였다. 최적의 액체배지조건을 확인하고자 IBA의 농도는 0.5 mg/L로 첨가하고 sucrose의 농도를 달리하여 실험한 결과 sucrose는 30 g/L 첨가 되었을 때 가장 높은 생중량과 건중량을 나타냈다. 액체배양된 여우구슬의 부정근을 각각 MS, 1/2MS, 1/3MS배지에 30 g/L sucrose, 0.5 mg/L IBA가 첨가된 5 L 용량의 생물반응기에 4주간 배양한 결과 1/2MS 배지에서 양호한 생장을 보였다. 본 실험에서는 여우구슬의 종자발아체를 이용하여 부정근의 유도 및 증식조건에 필요한 기내배양조건과 2차적으로 유도된 부정근을 이용하여 플라스크와 생물반응기 배양을 통한 효율적인 증식조건을 확립하였다.

In vitro proliferation system was achieved by using nodal segment excised from greenhouse grown juvenile stock plants of Alnus japonica. Stem explants were cultured on MS medium supplemented with different plant growth regulators of cytokinin and/or their combinations. The most effective cytokinin source was the combination of zeatin 2.0 mg/L and TDZ 0.05 mg/L producing the average number of shoots (16.8 ± 3.6). In addition, healthy roots were formed after small clumps of shoots were transferred to half strength of MS medium containing IBA 0.02 mg/L with optimal rooting capacity. Soil acclimatization was successfully conducted in cell tray containing artificially mixed soil with 92 % survival rate.

The efficiency of in vitro regeneration of four clones of Populus euramericana, Canada blanc, Eugenii, I-45/51, and Wisconsin #5, was examined. Cytokinins and the combinations with auxins affected the rate of regeneration from the explants of root segments, stem internodes, and leaf discs. Overall, BA and the combination with auxins were effective in root segments and leaf discs of the Canada blanc clone, whereas zeatin and the combination with auxins were important in stem internodes of the Wisconsin #5 clone. The highest number of shoots averaging 17.6 ± 0.47 from root segments in the Canada blanc clone,18.2 ± 3.0 from stem internodes in the Wisconsin #5 clone, and 17.8 ± 1.92 from leaf discs in the Canada blanc clone were obtained with 2.0 mg/1 BA, 2.0 mg/l zeatin combined with 0.2 mg/l IAA, and 0.5 mg/l BA combined with 0.05 mg/l 2,4-D, respectively. In particular, the addition of 2,4-D into cytokinin medium promoted shoot proliferation.

The effects of basal media, sucrose and phytohormone concentrations, and gelling agent combinations on in vitro frond proliferation of Lemna gibba G3 and 24 additional Lemna gibba strains were examined. Frond proliferation was equivalent on Schenk and Hidebrand. Murashige and Skoog. Nitsch and Nitsch, and Gamborg's B5 media and poor on murashige and Skoog medium in the absence of benzyladenine. With the addition of benzyladenine, Schenk and Hildebrand and Gamborg's B5 Were superior and equivalent. The addition of benzuyladenine increased equally frond proliferation at either 1 or 10μM, however at 10μM fronds were severely curled or fused. Benzyladenine and thidiazuron suppressed root growth but kinetin was found to greatly enhance root growth. Gibberellic acid inhibited frond proliferation. Frond proliferation was significantly different on the four sucrose concentrations of 0, 1, 3, and 5% Among them, 3% sucrose was found to be superior. The reduced frond size observed in cultures grown on 8% sucrose could be explained by showing medium osmotic potential in excess of frond water potential. Gell agents also varied significantly in their ability to promote frond proliferation with 0.25% Gelrite or a mixture of 0.15% Gelrite and 0.4% agar. Proliferation of 25 Lemna gibba strains on medium neat optimal for Lemna gibba G3 showed a six-fold variation across strains with Lemna gobba G3 placing in the top 5 fastest proliferating strains.