• 한국습지학회지
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• 제20권1호
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• pp.27-34
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• 2018

본 연구에서는 멸종위기식물인 큰바늘꽃(Epilobium hirsutum L.)의 효과적인 보전 및 복원을 위한 기초자료를 얻기 위해 토양의 수분함량과 유기물함량이 번식계절과 생리 반응에 어떠한 영향을 주는지에 대하여 알아보았다. 큰바늘꽃은 다년생식물이지만 모든 구배에서 한 해에 생식생장을 하였다. 꽃봉오리, 꽃 그리고 열매주머니는 수분구배와 영양소구배에서 각각 높은 수분 조건과 높은 유기물 조건에서 가장 이른 시기에 성숙하였다. 그리고 꽃 수와 열매주머니 수는 높은 수분 조건과 높은 유기물 조건에서 더 빨리 증가하였다. 엽록소 함량은 수분구배에서 높은 중간 수분 조건과 높은 수분 조건에서 가장 많았고, 영양소구배에서는 차이가 없었다. 최소엽록소형광 값은 수분구배와 영양소구배 모두 차이가 없었고, 최대엽록소형광 값은 높은 수분 조건과 높은 유기물 조건에서 가장 높았다. 광계 II의 광화학적 효율 값은 모든 수분구배에서 0.75로 차이가 없었고, 영양소구배에서의 경우 높은 유기물 조건에서 0.78로 가장 높았다. 큰바늘꽃은 수분이 증가할수록 엽록소 함량이 많아지고, 유기물이 증가할수록 Fv/Fm 값이 높아졌다. 이상의 연구결과는 토양의 충분한 수분과 유기물 함량은 큰바늘꽃의 번식계절을 앞당겨 주고 생식생장을 촉진한다는 것을 보여준다. 추후 멸종위기종인 큰바늘꽃의 개체군 유지와 서식지를 관리하는데 중요한 정보가 될 것으로 판단된다.

• Clinical and Experimental Reproductive Medicine
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• 제33권1호
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• pp.25-33
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• 2006

목 적: 본 연구에서는 착상전 생쥐 배아에서 분리한 할구를 이용하여 배아줄기세포주를 확립하고 그 효용성과 특성을 살펴보고자 하였다. 연구방법: 생쥐 (C57BL/6J)의 2- 또는 4-세포기 배아에서 투명대를 제거하고 할구를 분리하여 지지세포와 공동배양한 후 할구로부터 형성된 내세포괴를 분리하여 계대배양을 실시하였다. 계대배양 중인 세포주의 특성을 확인하기 위해 alkaline phosphatase 활성도와 표지 인자 및 관련 유전자 발현을 세포면역화학적 염색과 RT-PCR 방법으로 살펴보았다. 또한, 계대배양 중인 배아줄기 세포주의 염색체 분석을 실시하였다. 결 과: 전체적으로 2-세포기에서 분리한 할구와 4-세포기에서 분리한 할구에서 각각 3.0% (1/33)와 4.0% (1/25)의 효율로 배아줄기세포주를 확립할 수 있었다. 이는 4-세포기의 배아를 사용하였을 때의 16.7% (5/30)에 비해 현저하게 낮았다. 분리된 할구로부터 확립된 배아줄기세포주에서 SSEA-l 과 Oct-4의 발현을 관찰하였고, 이들에서 분화된 배아체에서 삼배엽성 분화 관련 유전자들의 발현도 확인할 수 있었다. 결 론: 본 연구에서는 동물모델을 이용하여 착상전 초기 배아에서 분리한 할구를 이용하여 배이줄기세포를 확립할 수 있음을 확인하였다. 지속적인 관련 연구를 통해 인간의 체외수정 및 배아이식술에서 배아의 파괴 또는 발생 능력에 손상을 주지 않고 새로운 인간 배아줄기세포주를 생산할 수 있는 방법을 개발하고 실용화할 수 있을 것으로 사료된다.

• 한국수정란이식학회지
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• 제29권1호
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• pp.7-12
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• 2014

본 연구는 한우 번식 기록이 잘 유지되고 있는 4개 농장의 2007년 1월부터 2010년 10월까지의 번식 자료 수집하여 분석하였다. 수태 당 평균 수정 횟수와 평균 공태일은 A농장 $1.7{\pm}0.1$회와 $77.4{\pm}4.8$일, B농장 $1.5{\pm}0.1$회와 $150.8{\pm}11.2$일, C농장 $1.5{\pm}0.1$회와 $90.4{\pm}4.5$일, D농장 $1.4{\pm}0.1$회와 $71.4{\pm}2.5$일이었다. 호르몬으로 발정을 유도하는 D농장을 제외한 3개 농장 531두의 번식 기록으로 분만 후 첫 수정 시기에 따른 평균 수정 횟수와 평균 공태일을 분석한 결과, 총 5개의 수정 시기에 따른 수정 횟수는 30일 이전 첫 수정이 $2.1{\pm}0.2$회로 31일 이후 첫 수정보다 유의적으로 높았다. 번식 장애우 58두에 2가지 배란 동기화법을 사용하여 수태율을 확인해 본 결과, Ovsynsh 법은 55.2%의 수태율을, CIDR-based TAI 법은 65.5%의 수태율을 나타냈다. 농장의 번식률을 높이기 위해서는 정확한 번식 기록 작성, 발정 관찰, 수정 후 임신 감정, 번식 기관 검진, 번식률을 고려한 첫 수정 시기 수정 등이 필요하다.

• 대한수의학회지
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• 제34권1호
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• pp.181-188
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• 1994

In 60 dairy cows with inactive ovaries, follicular cyst, luteal cyst, persistent corpus luteum and silent heat as diagnosed by rectal palpation, and those that had not resumed ovarian cycles until 60 days postpartum, progesterone concentrations for differential diagnosis of reproductive disorders were measured and were compared in matched plasma, skim milk and milk fat samples at 10 days interval. The incidence rate of reproductive disorders were as follows; inactive ovaries 20(33.3%), silent heat 11(18.3%), follicular cyst 7(11.7%), luteal cyst 7(11.7%), persistent corpus luteum 7(11.7%), pyometra 4(6.7%), vaginitis 2(3.3%), cystic corpus luteum 1(1.7%), and endometritis 1(1.7%), respectively. Cows having a progesterone concentration in plasma and skim milk < 1.0 ng/ml, and in milk fat < 80.0 ng/ml were considered to have inactive ovaries or follicular cyst. Those with concentrations in plasma and skim milk ${\geq}1.0ng/ml$, and in milk fat ${\geq}80.0ng/ml$ were regarded as the cases of luteal cyst or persistent corpus luteum. Progesterone concentrations in above cows did not differ significantly between the time of initial determination and the 10 days after initial determination. But progesterone concentrations in cows with silent heat did differ significantly between the time of initial determination and the 10 days after initial determination(P<0.05). The accuracy of rectal palpation for making a differential diagnosis of ovarian dysfunction, as defined on basis of progesterone concentrations, were as follows; follicular cyst 55.6%, luteal cyst 50.0%, inactive ovaries 90.5% and persistent corpus luteum 60.0%, respectively. It may be concluded that progesterone determinations at 10 days interval is practical as an aid to diagnosing ovarian dysfunction, particularly follicular cyst and luteal cyst.

• 동물자원연구
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• 제29권4호
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• pp.158-165
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• 2018

본 연구는 Canada에서 수입한 동결정액으로 인공수정된 모돈의 번식성적을 조사하여 동결정액을 이용한 인공수정(동결정액 인공수정)의 효율을 개선할 수 있는 통찰력을 얻기 위해 수행되었다. 이를 위해 A(GGP) 농장에서 2016년 5월과 9월 사이 총 626회의 동결정액 인공수정과 B(GGP) 농장에서 2015년부터 2017년 기간 동안 수행한 총 2,429 동결정액 인공수정의 결과 기록을 분석하였다. A종 돈장에서 동결정액을 썼을 때 총산자수 및 실산자수는 9월 보다 5월에 높았다(p<0.05). B종돈장의 분만율, 총산자수 및 실산자수는 조사연도 간에 차이가 없었다. A종돈장 결과와 B종돈장 결과를 통합하여 분석했을 때, 분만율은 A종돈장이 높았으나(p<0.01) 총산자수와 실산자수는 두 종돈장간에 차이가 없었고, 동결정액 인공수정을 했을 때가 액상정액 인공수정을 했을 때보다 낮았다(동결정액:액상정액 인공수정에 대한 총산자수와 실산자수는 각각 $10.9{\pm}0.3:13.4{\pm}0.1$두 및 $10.0{\pm}0.3:12.0{\pm}0.1$두 이었음; p<0.01). 결론적으로, 이상의 결과는 동결정액 인공수정이 액상정액 인공수정보다 번식 효율이 낮긴 하지만 연중 적절한 시기에 동결정액 인공수정을 실시하면 번식 효율이 증가될 수도 있음을 시사한다.

• 한국수정란이식학회지
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• 제30권4호
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• pp.345-350
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• 2015

In the last 10 years, porcine somatic cell nuclear transfer to generate transgenic pig has been performed tremendous development with introduction and knockout of many genes. However, efficiency of porcine somatic cell nuclear transfer is still low and embryo transfer (ET) is one of important step for production efficiency. In porcine ET for production of transgenic cloned pig, we can consider many of points to increase production rates. In respect of seasonality and weather, porcine ET usually is not performed in summer and winter. Cloned transgenic embryos must be transferred into reproductive tracts of recipients where embryos are located after natural fertilization with similar estrous cycle. If cloned embryos with 2~4 cell stage are transferred, they must be transferred into oviducts in periovulatory stage. Number and deposition sites of transferred cloned embryos are important. And we must compare the methods of ET between surgical and non-surgical ones in respect of production efficiency. Sow recipients after natural estrus is most preferred recipients however its cost is must be considered. Here we will review many of current studies about porcine embryo transfer to increase production efficiency of transgenic pigs and strategies for further studies.

• Reproductive and Developmental Biology
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• 제33권2호
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• pp.113-117
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• 2009

Animals produced by somatic cell nuclear transfer (SCNT) using genetically modified cells are almost always transgenic, implying that this method is more efficient than the traditional pronuclear microinjection method. Most somatic cells for SCNT in animals are fetus-derived primary cells and successful gene integration in somatic cells will depend on transfection condition. The objective of this study is to evaluate the efficiency of electroporation (Microporator) and liposome reagents (F-6, F-HD, W-EX, W-Q, W-M) for tissue-type plasminogen activator (tPA) gene transfection and to estimate the overall efficiency of transfection of Korean native pig fetal fibroblast cells (KNPFF). Electroporation showed significantly higher transfection efficiency than liposome reagents with regard to the transfection of in vitro cultures in the early stages of development (41.7% with Microporator vs. 18.3% with F-6, 20.0% with F-HD 18.5% with W-EX, 5.0% with W-M and 6.3% W-Q,). Colonies identified as tPA-positives were treated once more with G418 for 10 to 14 days and growing colonies were selected again. When the cells of newly selected colonies were subjected to single-cell PCR, reselection of colonies following second round of G418 selection increased the rate of transgene integration per each colony. These results suggest that transfection with electroporation is the most efficient and the second rounds of G418 selection may be an effective method for transfection of porcine fetal fibroblast cells.

• Reproductive and Developmental Biology
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• 제30권4호
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• pp.279-285
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• 2006

Vitrification has been suggested to be an effective method for the cryopreservation of human ES cells. However, the efficiency of vitrification with different vehicles remains a matter of ongoing controversy. The objective of this study was to assess the efficiency of cryopreservation in human ES cells by vitrification using different vehicles. A human ES cell line and a variety of vehicles, including micro-droplet (MD), open-pulled straw (OPS) and electron microscopic grid (EM-grid), were employed in an attempt to assess vitrification efficiency. In order to evaluate the survivability and the undifferentiated state of the post-vitrified human ES cells, we conducted alkaline phosphatase staining and characterization via both RT-PCR and immunofluorescence assays. The survival rates of the post-vitrified human ES cells using MD, OPS and EM-grid were determined to be 61.5%, 66.6% and 53.8%, respectively. There also exist significant differences between slow-freezing and vitrification (p<0.01). However, no significant differences were detected between the vehicle types. Finally, the pluripotency of human ES cells after thawing was verified by teratoma formation. Cryopreservation using vitrification is more effective than slow-freezing, and the efficiency of vehicles proved effective with regard to the preservation of human ES cells.

• Reproductive and Developmental Biology
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• 제35권4호
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• pp.449-455
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• 2011

Induced pluripotent stem (iPS) cells have been generated from mouse and human somatic cells by etopic expression of transcription factors. iPS cells are indistinguishable from ES cells in terms of morphology and stem cell marker expression. Moreover, mouse iPS cells give rise to chimeric mice that are competent for germline transmission. However, mice derived from iPS cells often develop tumors. Furthermore, the low efficiency of iPS cell generation is a big disadvantage for mechanistic studies. Nonviral plasmid.based vectors are free of many of the drawbacks that constrain viral vectors. The histone deacetylase inhibitor valproic acid (VPA) has been shown to improve the efficiency of mouse and human iPS cell generation, and vitamin C (Vc) accelerates gene expression changes and establishment of the fully reprogrammed state. The MEK inhibitor PD0325901 (Stemgent) has been shown to increase the efficiency of the reprogramming of human primary fibroblasts into iPS cells. In this report, we described the generation of mouse iPS cells devoid of exogenous DNA by the simple transient transfection of a nonviral vector carrying 2A-peptide-linked reprogramming factors. We used VPA, Vc, and the MEK inhibitor PD0325901 to increase the reprogramming efficiency. The reprogrammed somatic cells expressed pluripotency markers and formed EBs.

• Reproductive and Developmental Biology
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• 제28권1호
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• pp.7-12
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• 2004

This study was performed to establish the effective culture condition for the establishment of clonal lines from porcine fetal fibroblast cells. Fibroblasts derived from a pig fetus (Day 50) were cultured and passaged two times before use. A single cell was seeded in 96-well plates, cultured in medium supplemented with different concentrations of FBS, catalase or $\beta$-mercaptoethanol ($\beta$ME), and classified by cell size and morphology. Cells were passaged two times into 4-well dish before freezing. The establishment efficiencies were not different among different concentrations of FBS (0.3 to 5.1%). However, population doubling time (PDT) was significantly decreased by increasing the FBS concentration (P＜0.05). The establishment efficiency of $\beta$ME-added group (10.4%) was significantly higher than those of catalase-added and control groups (3.5%, and 3.5%, respectively, p＜0.05), and PDT was significantly decreased (23.6 vs 28.1, and 25.5 h, respectively, p＜0.05). However, catalase did not show a positive effect on the establishment efficiency. Cell size and morphology did not affect the establishment efficiency and PDT of clonal lines. The result of present study shows that the establishment efficiency of clonal cell lines can be enhanced by the culture in media supplemented with 30% FBS and $\beta$ME.