Trandgenic Mouse Embryo를 이용한 Human HoxA 유전자의 조절부위 분석과 전후축 형태형성(Anterior-Posterior Axial Pattern Formation)에 미치는 영향

장승익 충남대학교 1995 국내석사

The Hox genes are known to be involved in the anterior- posterior axial pattern formation through differential expression during early embryogenesis in both invertebrate and vertebrate. They are organized in the cluster(s), and each cluster is composed of 13 paralogous group. The localization of the gene in the cluster is colinear with the expression domain along the anterior-posterior axis as well as with the expression time. The regulatory element(s) which govern(s) the expression pattern had been analysed in mice using transgenic mice system. However nothing has been known in human. In order to investigate the function of human homolog of position specific element of mouse Hoxa-7, a transgene was constructed. It contains 1. 1kb human DNA(HCR)-a homolog to the intergenic region between Hoxa-7 and 9, which directs the position specific expression of Hoxa-7-, tk promoter, LacZ(β-galactosidase) gene as a reporter and polyadenylation signal of SV40 large T antigen. It was injected into the mice embryos, and the resulting transgenic embryos were analysed through PCR as well as genomic Southern blotting with placenta DHA. The results obtained through the experiments were summarized as follows; 1. The plasmid, pHJ-1, contains the human specific DNA fragment(HCR 1. 1 Kb) which is a homolog to the mouse Hoxa-7 control region. 2. HJ-1 tranagene [HCR 1. 1kb-tk promoter-LacZ gene-poly adenylation signal of SV40 large T antigen] was injected into the mice fertilized eggs, and 20 embryos were obtained. 3. The transgenic embryos were analysed through PCR as well as genomic Southern blotting with placenta DNA, and 2 embryos were found to be transgenic. 4. One out of two transgenic embryos expressed transgene when stained with A-gal. The expression pattern was in analogy to that of the mouse Hoxa-7 control region, showing spatially restricted expression pattern. Since the expression of LacZ(β-galactosidase) is regulated by the ustream human HCR sequence, it implies that the HCR is the plausible position specific regulatory el ement of human.

유전자 편집 기술의 윤리·법·사회 문제 해결방안

이수빈 연세대학교 대학원 2019 국내석사

Genome editing technology is likely to evolve into a technology that enables fundamental prevention and treatment of genetic diseases. Recently, genome editing technology has been expanded to include research on human embryos. However, a growing concern exists regarding the use of genome editing technology on human embryo that can cause permanent genetic changes to future generations. Moreover, international discussions continue on conducting this research. Therefore, this study is aimed at exploring the recognition of ethics, legal, and social issues in genome editing and proposing a solution for our country using the comparison and analysis of country-specific regulations through an in-depth interview. As a result, the following ethical, legal, and social issues in genome editing are determined. First, the safety in genome editing technology is low, and the possibility of technology abuse is high. Second, genome editing technology applied to human embryos is expected to have a high potential for embryonic rights to life and humanity. Third, the use of genome editing technology, which requires high cost, will increase the possibility of a social gap. Fourth, international and domestic guidelines on the regulation of genome editing research are insufficient. Comparison of the country-specific regulations has resulted in the establishment of laws or guidelines that specify genome editing research using human embryo in most countries. Furthermore, these countries permit human embryo genome editing for limited research purposes only and prohibit its use for the purposes of clinical research. As a solution for ethical, legal, and social issues in genome editing, a limited approach to prevent the misuse of technology and continuous research for confirming its safety are necessary. Application of genome editing technology on human embryo should be allowed for basic research only, but using it for the purpose of pregnancy and childbirth should be banned. Moreover, a discussion on the boundary between treatment and enhancement to prevent the possibility of human variability is needed. It should also seek to reduce costs through government subsidies or private insurance to alleviate social disparities and to mitigate the burden of state finances. International cooperation has caused the establishment of international guidelines for genome editing research regulations and the need to amend the Bioethics and Safety Act. Genome editing is controversial in terms of morality, ethics, and science; however, the possibility of eradicating diseases, which is a long-standing human problem, is highly expected. We need ongoing discussions and research on the appropriate use and regulation of genome editing. It is hoped that experts' recognition of the ethical, legal, and social issues in genome editing and the comparison and analysis of the country-specific regulations will serve as a solution to help establish the legality and system of genome editing in South Korea. 유전자 편집 기술은 유전질환의 근본적 예방 및 치료를 가능하게 하는 기술로 발전될 가능성이 높다. 최근 유전자 편집 기술은 인간 배아를 대상으로 하는 연구로 확대되어 가는 추세이다. 그러나 인간 배아에 적용하는 유전자 편집 기술은 다음 세대에 영구적인 유전자 변화를 줄 수 있다는 우려가 높아 연구를 수행함에 있어 국제적인 논의가 계속되고 있다. 이에 본 연구는 국내·외 전문가들을 대상으로 심층면접 연구를 시행하여 유전자 편집 기술의 윤리·법·사회 문제에 대한 인식을 확인하고, 국가별 규제의 비교·분석을 토대로 우리나라의 해결방안을 제시하였다. 연구 결과, 유전자 편집 기술의 윤리·법·사회 문제는 첫째, 유전자 편집 기술은 기술의 안전성이 낮고, 기술의 오남용 가능성이 높다. 둘째, 인간 배아에 적용하는 유전자 편집 기술은 배아의 생명권 문제와 인류의 변종 가능성이 높을 것으로 예상된다. 셋째, 고가의 비용이 요구되는 유전자 편집 기술의 활용으로 사회적 격차 발생 가능성이 높아질 것이다. 넷째, 유전자 편집 연구의 규제에 대한 국제 및 국내 가이드라인이 미흡한 상황이다.국가별 규제 현황을 비교한 결과, 대부분의 국가에서 인간 배아 유전자 편집 연구를 특정 하는 법률 또는 가이드라인이 존재하였다. 그리고 제한된 연구 목적으로 인간 배아 유전자 편집을 허용하고 있는 국가들 모두 임상 연구 목적은 금지한다는 규정을 함께 명시하고 있음을 발견하였다. 유전자 편집 기술의 윤리·법·사회 문제 해결방안으로는 기술의 안전성 확인을 위한 지속적 연구와 기술의 오남용 방지를 위한 제한적 접근이 필요하다. 인간 배아 유전자 편집 연구는 기초연구는 허용하되 임신 및 출산 목적은 금지되어야 하며, 인류의 변종 가능성 방지를 위한 치료와 강화의 경계 논의가 필요하다. 또한 사회적 격차 해소를 위한 정부지원 또는 개인보험을 통한 비용 절감과 국가재정부담을 완화하기 위한 제도를 모색해야 한다. 국제적 공조에 의하여 유전자 편집 연구 규제에 대한 국제 가이드라인 마련과 현행 생명윤리법 개정 필요성이 제시되었다. 유전자 편집 기술은 과학적, 윤리적, 법적, 사회적, 종교적 측면에서 여러 가지 논란이 일고 있지만, 인류의 오랜 문제인 질병 퇴치에 대한 가능성이 기대된다. 따라서 인류 전체의 보건 향상을 위하여 기술을 현명하게 활용하기 위한 지속적인 논의와 연구가 필요하다. 이번 연구에서 파악된 유전자 편집 기술의 윤리·법·사회 문제에 대한 전문가들의 인식과 국가별 규제의 비교·분석에 따른 해결방안이 향후 우리나라 유전자 편집 기술의 법·제도 방안 수립에 도움이 되기를 기대한다.

헌법상 배아의 인간존엄성과 생명권에 관한 연구

김용범 전북대학교 대학원 2010 국내석사

Recently, Researches on human embryos are in the spotlight as a last chance to treat the patients of incurable diseases. As a result of the remarkable developments in biotechnology, it has become an important issue whether an embryo should be afforded to human dignity and the right to life. The problems about human embryo researches are argued in several fields. These problems are going to be solved by legal system, especially in the constitutional methods. And the issues are whether biotechnology impinges upon fundamental rights such as human dignity and the right to life. Finally, the main issues are related with following questions : whether human embryos can be regarded as human beings, if how what human embryos can be regarded, and how research on embryos could be protected by constitution. Before, the constitutional discussion, the question had been discussed what status can be recognize to human embryos as preliminary to human beings. And I propose 'humanity', 'independence', and 'identity' as the conception attributes to human dignity. In the constitution 'human dignity', if the best value that should be guaranteed unconditionally, so this can not be examined stage by stage. In relation to the right to life, 'life' which is comprehended embryologically begins after insemination in aspect of continuity. Because the restriction of 'the right to life' can be justified on a particular case, 'the right to life' is a relative right. In criminal law, prescribed punishments about infringement on life are relative. Therefore, it means that the national protection obligation of the life is graded. The study has also examined the legislation procedures on life science and explained the necessity of legislation on embryonic research through significance and principal contents of the current 'Bioethics and Safety Act'. And I have presented the legislation on human embryo research in cases of the United States, Germany, the United Kingdom of Great Britain, Japan, and the trends in international organizations like UN, UNESCO, WHO. With the development of science and technology, mankind has been in close connection with scientific civilization. Particularly because of radical advances in biotechnology, human beings become to believe that of an incurable disease or new organ generation which had been regarded impossible becomes realistic and feasible. Consequently, biotechnology which has been regarded as beneficial is now being warned of its dangerousness and the necessity to put a restrict on biotechnology practices have been suggested. It`s difficult to draw a definite conclusion because there is a gap between research of biotechnology and ethical and religious sense of value. Therefore I have suggested an outline to solve an argument through a legal discussion. Of course, all the problems related to this field can not be solved out completely. keywords : Human embryo, Human Dignity, Bioethics, The Right to The Life, Bioethics and Safety Act Student ID Number : 200750689

Modeling Human Epiblast Morphogenesis

Resto, Agnes M University of Michigan ProQuest Dissertations & Th 2023 해외박사(DDOD)

The development of the human embryo is arguably the most complex process that we could care to study. In this process, the developing embryo must undergo proliferation, reorganization, lineage diversification, and dozens of cell fate specification events. During this time, a myriad of events are happening in parallel at the cell level, each one setting the foundation for the emergence of increasingly complex tissues of increasingly complex function. Understanding the mechanisms guiding these processes is pivotal not only for embryogenesis-related applications in fertility and development, but also for regenerative medicine applications such as the development of organ replacements.In this dissertation, I propose an integrative approach to the study of morphogenesis and patterning, specifically in the context of stem cell-based models of human development. Firstly, I present a novel machine learning-assisted imaging pipeline that permits the careful characterization of cell-level events occurring in our in vitro model of epiblast cyst morphogenesis. Secondly, I present a novel agent-based model (ABM)-genetic algorithm (GA) framework for the generation of models of morphogenesis. The framework was first tested to determine its ability to generate structures of desired patterns. It was then applied for the generation of models that plausibly capture mechanisms at work during epiblast cyst morphogenesis and symmetry breaking. With preliminary in silico experiments, I showed that the framework was able to output models that partially captured the effect of initial cell number on final cyst composition. I further showed that correct structure formation was heavily impacted by just a few model parameters. Combined with in vitro experimentation, these tools have the potential to shed light into the mechanisms guiding growth, movement, and cell fate specification in in vitro models of human development.

배자기의 원시적혈구모세포에 관한 형태계측학적 연구

주영식 중앙대학교 1991 국내박사

인간 식도의 조직발생

박인규 연세대학교 대학원 2002 국내석사

선천성 기형의 기전 이해, 진단 및 적절한 처치를 위해서는 정상 발생에 관한 지식이 필수적이다. 이 연구에서는 발생 4주부터 8주까지의 인간배자 11예와 발생 12 - 30주 사이의 태아 14예 등 25예에서 식도의 정상조직 분화과정을 관찰하여 다음과 같은 결과를 얻었다. 식도는 발생 4주말에 전장관의 팽창된 부분으로 처음 구별되었으며, 식도의 앞쪽에는 후두기관관과 폐싹이 발달해 있었다. 식도를 둘러싸는 미분화 중간엽은 발생 6주말(발생 17기)에 분명하게 구분되었다. 식도의 상피는 발생 초기에는 2 - 3층의 원주세포로 구성되어 있었다. 이들 상피세포는 계속 증식하여 발생 7주 및 8주 초에 상피가 5 - 6층으로 두꺼워졌고 동시에 식도의 관강이 매우 좁아졌으며, 상피 속에 다수의 소포들이 형성되었다. 발생 8주말부터 소포들이 없어지면서 다시 하나의 큰 관강을 형성하였다. 발생 12주에는 식도의 점막에서 4개의 일차주름이 처음 관찰되었다. 발생 12주부터 20주까지는 상피의 표층에 섬모화 원주세포가 계속 증가했으며, 발생 20주 이후 표층세포가 납작한 편평상피로 교체되기 시작해 발생 30주에는 식도의 상피가 성인에서와 같은 중층편평상피의 특징을 나타내었다. 이러한 상피세포의 변화는 식도 중간부부터 시작되어 위, 아래로 진행되었다. 점막근육판은 발생 12주에 식도의 하부에서 처음 발달하였고, 고유판은 발생 16주부터 구분이 가능하였다. 점막하층과 윤주근층의 원기는 발생 5주말에 처음 구분할 수 있었다. 발생 8주에 윤주근층은 상당히 성숙한 상태였고, 종주근층이 관찰되기 시작하였다. 발생 12주에는 종주근층도 뚜렷해졌다. 근육층은 식도의 하부로부터 발생하기 시작하였다. The knowledge about normal development is essential for understanding and managing the congenital anomalies. In this study, normal esophguses of consequent 11 human embryos and human fetus were inspected under light microscopy. The esophagus was demarcated from other portions of the foregut at the end of the 4th week. Initially, esophageal epithelium was composed with 2-3 layers of columnar cells. The epithelium undergoes progressive proliferation and became thickened. The Lumen became narrowed but not occluded. At the 8th week vacuoles appeared in epithelium. From the end of the 8th week, as vacuoles disappeared single large lumen was restored. The lumen of esophagus has assumed the form of a Greek cross by four primary folds at the 12th week. From that time ciliated cells appeared on superficial layer of the epithelium. Ciliated epithelium covered almost area of esophagus at the 20th week. From 20th week ciliated epithelium was replaced by squamous epithelium. At 30th week, adult type non-keratinized stratified squamous epithelium was observed. The epithelial change was started from the midesophagus. The muscularis mucosa was developed from the 12th week. The lamina propria was observed at the 16th week. The primordia of submucosa and muscle layers were distinguishable at the end of the 5th week. The circular layer was well developed at the 8th week and the longitudinal layer was apparent at the 12th week. The separation process of esophagus and trachea is not fully understood. Further studies with human specimens and animal are needed to clarify normal developmental process and pathogenesis of congenital anomalies of the esophagus.

사람태아 간장의 적혈구조혈세포에 관한 핵 계측학적 연구

윤경준 중앙대학교 1988 국내박사

사람 배자기의 직장과 항문관의 형태형성에 관한 연구

홍정 연세대학교 1991 국내박사

인배자와 태아에서 측두하악관절의 발생에 관한 형태학적 연구

연용흠 연세대학교 1997 국내박사

정상치배 발생 과정과 법랑아세포종에서 Transforming growth factor-beta 발현에 관한 면역조직화학적 연구

성길현 원광대학교 1997 국내박사