• Title/Summary/Keyword: Fetal

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Effect of Fusion Condition on In Vitro Development of Caprine Cloned Oocytes with Nuclear Transfer (재래산양에 있어서 핵이식란의 융합조건이 융합 및 체외발달에 미치는 영향)

  • 박희성;김태숙;이윤희;정수영;이명열;홍승표;박준규;김충희;정장용
    • Reproductive and Developmental Biology
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    • v.28 no.2
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    • pp.127-132
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    • 2004
  • This study was conducted to examine the effects of electric stimulation conditions on in vitro developmental ability of caprine embryos after somatic cell nuclear transfer. Recipient oocytes were surgically collected after superovulation by using CIDR and FSH, PMSG, hCG and estrous synchronization in Korean native goats. The caprine ear cells were cultured in vitro in serum-starvation condition (TCM-l99 + 0.5% FBS) for 3 to 5 days of cell confluence. The zona pellucida of in vivo and in vitro matured oocytes were partially drilled using laser system. Single somatic cell was individually transferred into the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3M mannitol. After the electofusion, embryos were activated by electric stimulation or Ionomycin + 6-DMAP. Nuclear transfer embryos were cultured in mSOF medium supplemented with 0.8% BSA 6∼7 days at 39 , 5% $CO_2$, 5% $O_2$, 90% $N_2$. The fusion rate of donor cells was 60.4% and 40.3 % in ear cell and fetal fibroblast, and cleavage rate were 40.6% and 48.2%, respectively. No significant difference was found in the fusion and cleavage rate in different donor cells. Nuclear transferred oocytes were fused by electric pulses of 1.30∼1.40, 2.30∼2.39 and 2.40∼2.46 ㎸/cm. There was no significant difference among different electric pulses in fusion rates (26.7, 34.8 and 43.8%). The cleavage rate was higher (p<0.05) in 1.30∼1.40 ㎸/cm (82.9%) than 2.30∼2.39 ㎸/cm (43.8%) and 2.40∼2.46 ㎸/cm. (51.8%). The fusion rates of recipient oocyte source were 1st (43.5% and 23.6%), 2nd (55.7% and 39.2%) and 3rd (66.1% and 52.8%) in in vivo and in vitro oocytes. However, fusion ratee were significantly higher (p<0.05) in in vivo than in vitro oocyte. The cleavage rate of fused oocytes from in vivo and in vitro sources were 52.6% and 54.4%, respectively. No significant difference was found in the cleavage rate according to the recipient oocyte source. These results suggest that factors such as field pulse of electric stimulation and oocyte source could affect in vitro developmental ability of nuclear transplanted caprine oocytes.

Progesterone assays as an aid for improving reproductive efficiency in dairy cattle V. Plasma progesterone determination as applied to the differential diagnosis of reproductive disorders and judgement of treatment responses to PGF2α or GnRH treatment (Progesterone 농도측정(濃度測定)에 의한 유우(乳牛)의 번식효율증진(繁殖效率增進)에 관한 연구(硏究) V. 혈장(血漿) progesterone 농도측정(濃度測定)에 의한 무발정(無發情)의 감별진단(鑑別診斷) 및 PGF2α 또는 GnRH 치료효과(治療效果)의 판정(判定))

  • Kang, Byong-kyu;Choi, Han-sun;Son, Chang-ho;Oh, Ki-seok;Kang, Hyun-ku;Kim, Sam-ju;Kim, Hyek-jin;Kim, Nam-ki
    • Korean Journal of Veterinary Research
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    • v.35 no.3
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    • pp.603-613
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    • 1995
  • Plasma progesterone($P_4$) assay has been introduced to apply to the differential diagnosis of reproductive disorders and the monitoring of responses of ovarian dysfunction to $PGF_2{\alpha}$ or GnRH treatment in the 204 postpartum and postinsemination subestrus dairy cows. 1. The incidence rate of reproductive disorders in 204 subestrus cows, diagnosed by palpation per rectum and plasma $P_4$ determination using 'Two sample test'(Day 0+Day 10) were as follows; silent heat or error of estrus detection 110(53.9%), persistent corpus luteum 26(12.7%), follicular cyst 16(7.8%), inactive ovary 12(5.9%), luteal cyst 11(5.4%), granulosa cell tumor of ovary 1(0.5%), fetal mummification 1(0.5%), endometritis 15(7.4%) and pyometra 12(5.9%), respectively. 2. After the $PGF_2{\alpha}$ treatment to the 76 cows with silent heat or error of estrus detection, persistent corpus luteum, or luteal cyst, plasma $P_4$ concentrations at day 3 post treatment using 'Two sample test'(Day 0+Day 3) remained low(<1.0ng/ml) in all 76 cows. Therefore all 76 cows responded positively to $PGF_2{\alpha}$ treatment. Seventeen cows with follicular cyst or inactive ovary were treated with GnRH. All 7 cows with follicular cyst and 4 cows with inactive ovary remained high($${\geq_-}1.0ng/ml$$) a plasma $P_4$ concentrations at day 12 post treatment using 'Two sample test'(Day 0+Day 12), but 6 cows with inactive ovary remained low(<1.0ng/ml) a plasma $P_4$ concentrations. Therefore all 7 cows with follicular cyst and 4 cows with inactive ovary responded positively, but 6 cows with inactive ovary responded negatively to GnRH treatment. 3. The mean days from treatment to first service, number of cows conceived on first service(%), mean number of services per conception, mean days from initial treatment to conception, and mean number of cows conceived by 100 days post treatment(%) were 5.0 and 26.2 days, 45(59.2%) and 6(35.3%) cows, 1.5 and 1.7 services, 13.6 and 22.6 days, and 62(81.6%) and 9(52.9%) cows in group of $PGF_2{\alpha}$ and GnRH treatment, respectively. These results indicated that plasma $P_4$ assay was practical as an aid to diagnosing reproductive disorders and to monitoring responses of ovarian dysfunction to $PGF_2{\alpha}$ and GnRH treatment in subestrus cows.

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Review of 2011 Major Medical Decisions (2011년 주요 의료 판결 분석)

  • Yoo, Hyun-Jung;Seo, Young-Hyun;Lee, Jung-Sun;Lee, Dong-Pil
    • The Korean Society of Law and Medicine
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    • v.13 no.1
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    • pp.199-247
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    • 2012
  • According to the review and analysis of medical cases that are assigned to the Supreme Court and all local High Court in 2011 and that are presented in the media, it was found that the following categories were taken seriously, medical and pharmaceutical product liability, the third principle of trust between medical institutions, negligence and causation estimation, responsibility limit, the meaning of medical records and related judgment of disturbed substantiation, Oriental doctors' duties to explain the procedures, IMS events, whether one can claim for each medical care operated by non-physician health care institutions to the nonmedical domain in the National Health Insurance Corporation, and the basis of norms for each claim. In the cases related to medical pharmaceutical product liability, Supreme Court alleviated burden of proof for accidents with medical and pharmaceutical products prior to the practice of Product Liability Law and onset the point of negative prescription as the time of damage strikes to condition feasibility of the specific situation. In the cases related to the 3rd principle of trust between medical institutions, the Supreme Court refused to sentence the doctor who has trusted the judgment of the same third-party doctors the violations of the care duty. With respect to proof of a causal relationship and damages in a medical negligence case, the Supreme Court decided that it is unjust to deny negligence by the materials of causal relationship rejecting the original verdict and clarified that the causal relationship shall not deny the reasons to limit doctors' responsibilities. In order not put burden on patients with disadvantages in which medical records and the description of the practice or the most fundamental and important evidence to prove negligence and causation are being neglected, the Supreme Court admitted in the hospital's responsibility for the case of the neonate death of suffocation without properly listed fetal heart rate and uterine contraction monitor. On the other hand, the Seoul Western District Court has admitted alimony for altering and forging medical records. With respect to doctors' obligations to description, the Supreme Court decided that it is necessary to explain the foreseen risks by the combination of oriental and western medicines emphasizing the right of patient's self-determination. However, questions have arisen whether it is realistically feasible or not. In a case of an unlicensed doctor performing intramuscular stimulation treatment (IMS), the Supreme Court put off its decision if it was an unlicensed medical practice as to put limitation of eastern and western medical practices, but it declared that IMS practice was an acupuncture treatment therefore the plaintiff's conduct being an illegal act. In the future, clear judgment on this matter should be made. With respect to the claim of bills from non-physical health care institutions, the Supreme Court decided to void it for the implementation of the arrangement is contrary to the commitments made in the medical law and therefore, it is invalid to claim. In addition, contrast to the private healthcare professionals, who are subject to redemption according to the National Healthcare Insurance Law, the Seoul High Court explicitly confirmed that the non-professionals who receive the tort operating profit must return the unjust enrichment and have the liability for damages. As mentioned above, a relatively wide range of topics were discussed in medical field of 2011. In Korea's health care environment undergoing complex changes day by day, it is expected to see more diverse and in-depth discussions striding out to the development in the field of health care.

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Guidelines for the Management of Hydronephrosis Detected in the Perinatal Period (주산기에 발견된 수신증의 자연 경과와 치료 방침)

  • Chae Soo-Ho;Lee Ji-Hyuk;Jin Dong-Kyu;Park Kwan-Hyun;Paik Kyung-Hoon
    • Childhood Kidney Diseases
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    • v.10 no.1
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    • pp.33-39
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    • 2006
  • Purpose : Neonatal hydronephrosis is diagnosed with an incidence of 4.5-7% of pregnancies. Recently, early detection of neonatal hydroneprosis with antenatal ultrasonography has be-come possible. But consensus about its management has not been reached, especially concerning surgical intervention. The purpose of this study is to analyze the natural course of neonatal hydronephrosis and to determine the guideline of surgical intervention and follow up study. Methods : Sixty nine hydronephrotic kidneys were confirmed from April 2001 to April 2005. All cases were rechecked by ultrasonography once at least and had a minimum follow-up of 6 months. We classified the patients into 4 groups according to the anterior posterior pelvic diameter(APPD) on perinatal ultrasonography. Ultrasonography to measure the APPD diameter and Society for Fetal Urology(SFU) grade, $^{99m}Tc-MAG3$ scan were done to a set protocol. Pyeloplasty was peformed according to the protocol. Results : Pyeloplasty was not needed in most cases where the APPD was below 10 mm and where the SFU grade were less than grade 3. Many cases with APPD 10 mm above or SFU grade III above had undergone pyeloplasty. We found a correlation between obstruction grade on MAG3 scan and whether surgery was performed or not. Conclusion : If APPD is above 10 mm, SFU grade is above grade 3 or urinary tract obstruction is suspected by MAG3 scan, pyeloplasty must be considered. In cases where APPD is below 10mm and SFU grade is less than grade 3, we can observe the natural course of neonatal hydronephrosis with consecutive follow-up. (J Korean Soc Pediatr Nephrol 2006;10:33-39)

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The effect of tumor necrosis factor (TNF)-α to induce matrix metalloproteinase (MMPs) from the human dental pulp, gingival, and periodontal ligament cells (사람의 치수, 치은, 치주인대 세포에 tumor necrosis factor (TNF)-α로 자극 시 matrix metalloproteinase (MMPs)의 분비에 관한 연구)

  • Rhim, Eun-Mi;Park, Sang-Hyuk;Kim, Duck-Su;Kim, Sun-Young;Choi, Kyoung-Kyu;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.36 no.1
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    • pp.26-36
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    • 2011
  • Objectives: In the present study, three kinds of tissues cells (pulp, gingiva, and periodontal ligament) were investigated if those cells express MMP and TIMP when they were stimulated with neuropeptides (substance P, CGRP) or proinflammatory cytokine, TNF-$\alpha$. Materials and Methods: The cells cultured from human dental pulp (PF), gingiva (GF) and periodontal ligament were (PDLF) stimulated with Mock, SP, TNF-$\alpha$, and CGRP for 24 hrs and 48 hrs. for an RNase protection assay and Enzyme Linked Immunosorbent Assay. Cells (PF, GF and PDLF) seeded in 100 mm culture dish were stimulated with SP ($10^{-5}$, $10^{-8}\;M$) or only with medium (Mock stimulation) for 4hrs and for 24 hrs for RNase Protection Assay, and they were stimulated with CGRP ($10^{-5}\;M$) and TNF-$\alpha$(2 ng/mL) for 24 hrs and with various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for Rnase Protection Assay with a human MMP-1 probe set including MMP 1, 2, 8, 7, 8, 9, 12, and TIMP 2, 3. In addition, cells (PF, GF and PDLF) were stimulated with Mock and various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for 24 hrs and with TNF-$\alpha$(10 ng/mL) for 48 hrs, and the supernatents from the cells were collected for Enzyme Linked Immunosorbent Assay (ELISA) for MMP-1 and MMP-13. Results: The expression of MMPs in PF, GF, PDLF after stimulation with SP and CGRP were not changed compared with Mock stimulation for 4 hrs and 24 hrs. The expression of MMP-1, -12, -13 24 hrs after stimulation with TNF-$\alpha$ were upregulated, however the expression of TIMP-3 in PF, GF, PDLF after stimulation with TNF-$\alpha$ were downregulated. TNF-$\alpha$(2 ng/mL, 10 ng/mL, 100 ng/mL) increased MMP-1 and MMP-12 expression in PF dose dependently for 24 hrs. Conclusions: TNF-$\alpha$ in the area of inflammation may play an important role in regulating the remodeling of dentin, cementum, and alveolar bone.

Effects of Iron Overload during Pregnancy on Oxidative Stress in Maternal Rats (임신 쥐의 철 과잉섭취가 조직의 산화적 스트레스에 미치는 영향)

  • Park, Mi-Na;Lee, Yeon-Sook
    • Journal of Nutrition and Health
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    • v.44 no.1
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    • pp.5-15
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    • 2011
  • Although iron is an essential mineral, excess iron intake during pregnancy may increase oxidative stress in tissues. This study was conducted to investigate the effects of iron overload during pregnancy on iron status and oxidative stress in maternal rats. Ten week-old female Sprague-Dawley rats were mated with male rats. Non-pregnant (control) and pregnant rats were fed diets containing normal Fe (35 mg/kg diet), high Fe (350 mg/kg diet), or excess Fe (1,050 mg/kg diet) during pregnancy. Rats were sacrificed on pregnancy day 19. No significant difference in weight gain, diet intake, or litter size was observed according to iron intake levels. Furthermore, serum iron, hemoglobin, and hematocrit were not different among the rats administered the three levels of Fe both in the control and pregnant groups. However, the iron levels were lower in pregnant rats than those in the control. The liver and spleen iron contents increased significantly in the excess Fe group. An increase in liver ferritin levels with increasing iron intake was observed. Protein carbonyl content, as a marker of oxidative stress, increased significantly in liver with increasing iron intake but not malondialdehyde. Glutathione peroxidase activity in the liver of pregnant rats fed excess iron decreased significantly. Bcl-2 protein expression in the liver declined remarkably with increasing maternal iron intake in pregnant rats. Taken together, iron overload during pregnancy had little effect on hematology. However, the deposits of iron in the liver and the decline in antioxidant enzyme activity implied increased oxidative stress in tissues of the excess Fe group. These results suggest that excess iron intake during pregnancy increases oxidative stress in maternal tissues and may also affect fetal tissues.

Comparison of Ultrasound Image Quality using Edge Enhancement Mask (경계면 강조 마스크를 이용한 초음파 영상 화질 비교)

  • Jung-Min, Son;Jun-Haeng, Lee
    • Journal of the Korean Society of Radiology
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    • v.17 no.1
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    • pp.157-165
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    • 2023
  • Ultrasound imaging uses sound waves of frequencies to cause physical actions such as reflection, absorption, refraction, and transmission at the edge between different tissues. Improvement is needed because there is a lot of noise due to the characteristics of the data generated from the ultrasound equipment, and it is difficult to grasp the shape of the tissue to be actually observed because the edge is vague. The edge enhancement method is used as a method to solve the case where the edge surface looks clumped due to a decrease in image quality. In this paper, as a method to strengthen the interface, the quality improvement was confirmed by strengthening the interface, which is the high-frequency part, in each image using an unsharpening mask and high boost. The mask filtering used for each image was evaluated by measuring PSNR and SNR. Abdominal, head, heart, liver, kidney, breast, and fetal images were obtained from Philips epiq5g and affiniti70g and Alpinion E-cube 15 ultrasound equipment. The program used to implement the algorithm was implemented with MATLAB R2022a of MathWorks. The unsharpening and high-boost mask array size was set to 3*3, and the laplacian filter, a spatial filter used to create outline-enhanced images, was applied equally to both masks. ImageJ program was used for quantitative evaluation of image quality. As a result of applying the mask filter to various ultrasound images, the subjective image quality showed that the overall contour lines of the image were clearly visible when unsharpening and high-boost mask were applied to the original image. When comparing the quantitative image quality, the image quality of the image to which the unsharpening mask and the high boost mask were applied was evaluated higher than that of the original image. In the portal vein, head, gallbladder, and kidney images, the SNR, PSNR, RMSE and MAE of the image to which the high-boost mask was applied were measured to be high. Conversely, for images of the heart, breast, and fetus, SNR, PSNR, RMSE and MAE values were measured as images with the unsharpening mask applied. It is thought that using the optimal mask according to the image will help to improve the image quality, and the contour information was provided to improve the image quality.

Comparative Study of Compliance for Venous Thromboembolism Prophylaxis (American College of Chest Physicians Guideline and American College of Orthopedic Surgeons Guideline) in High Risk Patients with a Venous Thromboembolism (정맥혈전 색전증 고위험군에서 시행한 혈전색전 예방요법(American College of Chest Physicians 지침 및 American College of Orthopedic Surgeons 지침)의 준수율 비교 연구)

  • Suh, You-Sung;Nho, Jae-Hwi;Jang, Byung-Woong;Kang, Deokwon;Won, Sung-Hun
    • Journal of the Korean Orthopaedic Association
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    • v.54 no.4
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    • pp.317-326
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    • 2019
  • Purpose: To compare and analyze the rate of prevention of two venous thromboembolism prophylaxis guidelines in patients with artificial joint arthroplasty and hip joint fracture. Proper prophylaxis for preventing thromboembolism in orthopedic surgery is significant because of this fetal complication. Materials and Methods: This study compared and retrospective analyzed the rate of prevention using the medical records and radiographs of patients who underwent orthopedic surgery from March 2009 to February 2011 according to the American College of Chest Physicians (ACCP) guidelines and from March 2012 to February 2014 according to the American Academy of Orthopedic Surgeons (AAOS) guidelines. Results: The guidelines for venous thromboembolism prophylaxis have been applied to patients with artificial joint replacement and hip joint fracture, the compliance rate of the ACCP guidelines was 56.0% before surgery, 67.0% after surgery with chemical prophylaxis, and 80.5% with mechanical prophylaxis. In addition, the compliance rate of the AAOS guidelines was 74.1% with chemical prophylaxis, and 88.3% with mechanical prophylaxis, which was higher than the ACCP guidelines. The compliance rates of mechanical and chemical prophylaxis before and after surgery of the ACCP guidelines, and the compliance rate of mechanical and chemical prophylaxis of the AAOS guidelines were compared and analyzed. The results revealed statistical significance (p<0.05) before and after total knee replacement arthroplasty and hip joint fracture internal fixation and total high risk orthopedic surgery. Conclusion: Raising the compliance rate of prophylaxis of venous thromboembolism in high risk orthopedic surgery is necessary and people should follow the guidelines for a unified direction depending on which situation they are in.

Evaluation of Serum Insulin-Like Growth Factor(IGF)-I, Insulin-Like Growth Factor Binding Protein(IGFBP)-2 and IGFBP-3 Levels in Healthy Korean Children (정상 어린이에서 혈청 인슐린양 성장인자-I과 인슐린양 성장인자 결합단백-2 및 -3의 농도 분석)

  • Yang, Gi Hoon;Jung, Hye Lim;Kim, Deok Soo;Shim, Jae Won;Shim, Jung Yeon;Park, Moon Soo
    • Clinical and Experimental Pediatrics
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    • v.48 no.3
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    • pp.298-305
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    • 2005
  • Purpose : We performed this study to evaluate the mean serum levels of insulin-like growth factor (IGF)-I, insulin-like growth factor binding protein(IGFBP)-2 and IGFBP-3 in healthy Korean children according to age and sex. Methods : Ninety two healthy children, consisting of 42 boys and 50 girls, were classified into five groups according to age : neonate; infancy; early childhood; late childhood; and adolescence. We measured serum levels of IGF-I, IGFBP-2 and IGFBP-3 by enzyme-linked immunosorbent assay(ELISA) and analysed the serum levels according to sex and age group. Results : For boys, the mean serum levels of IGF-I(ng/mL) in neonate, infancy, early childhood, late childhood and adolescence were $41.1{\pm}3.6$, $70.9{\pm}33.7$, $103.5{\pm}97.2$, $89.8{\pm}46.5$ and $51.4{\pm}27.8$, respectively. Those of IGFBP-2(ng/mL) were $8.2{\pm}3.4$, $5.8{\pm}0.4$, $9.3{\pm}4.0$, $9.5{\pm}1.1$ and $7.0{\pm}0.5$, respectively. Those of IGFBP-3(ng/mL) were $559.2{\pm}215.2$, $1,333.3{\pm}692.5$, $2,254.6{\pm}1,513.8$, $2,447.1{\pm}1,464.2$, $1,533.6{\pm}807.4$, respectively. For girls, the mean serum levels of IGF-I(ng/mL) according to five age groups were $53.3{\pm}9.5$, $99.3{\pm}45.8$, $69.6{\pm}51.1$, $106.2{\pm}67.0$ and $145.1{\pm}127.8$, respectively. Those of IGFBP-2 (ng/mL) were $9.1{\pm}7.4$, $5.3{\pm}0.9$, $6.9{\pm}2.0$, $10.5{\pm}3.0$ and $7.9{\pm}1.3$, respectively. Those of IGFBP-3(ng/mL) were $858.2{\pm}433.4$, $1,834.8{\pm}851.3$, $1,404.3{\pm}570.2$, $2,203.5{\pm}899.4$ and $2,029.3{\pm}1,316.7$, respectively. There were significant positive correlations observed between IGF-I and IGFBP-3 levels(r=0.589, P=0.000). Conclusion : IGF-I and IGFBP-3 levels increased as children get older. The peak level of IGFBP-3 was observed in late childhood for both boys and girls, suggesting a current trend of children reaching peak growth velocity before adolescence. The IGFBP-2 level was higher in neonates compare to infancy, suggesting that IGFBP-2 is an important substance for fetal growth.

Preselection of Bovine Blastocysts Expressing Exogeneous Gene Following Microinjection (외래유전자를 주입한 소 수정란에서 형질전환가능 수정란의 선발)

  • 공일근
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.167-176
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    • 1997
  • This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

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