Direct Reprogramming of Human Dermal Fibroblasts Into Endothelial Cells Using ER71/ETV2

RATIONALE: Direct conversion or reprogramming of human postnatal cells into endothelial cells (ECs), bypassing stem or progenitor cell status, is crucial for regenerative medicine, cell therapy, and pathophysiological investigation but has remained largely unexplored. OBJECTIVE: We sought to directly reprogram human postnatal dermal fibroblasts to ECs with vasculogenic and endothelial transcription factors and determine their vascularizing and therapeutic potential. METHODS AND RESULTS: We utilized various combinations of 7 EC transcription factors to transduce human postnatal dermal fibroblasts and found that ER71/ETV2 (ETS variant 2) alone best induced endothelial features. KDR+ (kinase insert domain receptor) cells sorted at day 7 from ER71/ETV2-transduced human postnatal dermal fibroblasts showed less mature but enriched endothelial characteristics and thus were referred to as early reprogrammed ECs (rECs), and did not undergo maturation by further culture. After a period of several weeks' transgene-free culture followed by transient reinduction of ER71/ETV2, early rECs matured during 3 months of culture and showed reduced ETV2 expression, reaching a mature phenotype similar to postnatal human ECs. These were termed late rECs. While early rECs exhibited an immature phenotype, their implantation into ischemic hindlimbs induced enhanced recovery from ischemia. These 2 rECs showed clear capacity for contributing to new vessel formation through direct vascular incorporation in vivo. Paracrine or proangiogenic effects of implanted early rECs played a significant role in repairing hindlimb ischemia. CONCLUSIONS: This study for the first time demonstrates that ER71/ETV2 alone can directly reprogram human postnatal cells to functional, mature ECs after an intervening transgene-free period. These rECs could be valuable for cell therapy, personalized disease investigation, and exploration of the reprogramming process.ope

Generation of Human Pluripotent Stem Cell-derived Endothelial Cells and Their Therapeutic Utility

PURPOSE OF REVIEW: Human pluripotent stem cell-derived endothelial cells (hPSC-ECs) emerged as an important source of cells for cardiovascular regeneration. This review summarizes protocols for generating hPSC-ECs and provides an overview of the current state of the research in clinical application of hPSC-derived ECs. RECENT FINDINGS: Various systems were developed for differentiating hPSCs into the EC lineage. Stepwise two-dimensional systems are now well established, in which various growth factors, small molecules, and coating materials are used at specific developmental stages. Moreover, studies made significant advances in clinical applicability of hPSC-ECs by removing undefined components from the differentiation system, improving the differentiation efficiency, and proving their direct vascular incorporating effects, which contrast with adult stem cells and their therapeutic effects in vivo. Finally, by using biomaterial-mediated delivery, investigators improved the survival of hPSC-ECs to more than 10 months in ischemic tissues and described long-term behavior and safety of in vivo transplanted hPSC-ECs at the histological level. hPSC-derived ECs can be as a critical source of cells for treating advanced cardiovascular diseases. Over the past two decades, substantial improvement has been made in the differentiation systems and their clinical compatibility. In the near future, establishment of fully defined differentiation systems and proof of the advantages of biomaterial-mediated cell delivery, with some additional pre-clinical studies, will move this therapy into a vital option for treating those diseases that cannot be managed by currently available therapies.ope

Generation of Red Blood Cells from Human Pluripotent Stem Cells—An Update

Red blood cell (RBC) transfusion is a lifesaving medical procedure that can treat patients with anemia and hemoglobin disorders. However, the shortage of blood supply and risks of transfusion-transmitted infection and immune incompatibility present a challenge for transfusion. The in vitro generation of RBCs or erythrocytes holds great promise for transfusion medicine and novel cell-based therapies. While hematopoietic stem cells and progenitors derived from peripheral blood, cord blood, and bone marrow can give rise to erythrocytes, the use of human pluripotent stem cells (hPSCs) has also provided an important opportunity to obtain erythrocytes. These hPSCs include both human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). As hESCs carry ethical and political controversies, hiPSCs can be a more universal source for RBC generation. In this review, we first discuss the key concepts and mechanisms of erythropoiesis. Thereafter, we summarize different methodologies to differentiate hPSCs into erythrocytes with an emphasis on the key features of human definitive erythroid lineage cells. Finally, we address the current limitations and future directions of clinical applications using hiPSC-derived erythrocytes. © 2023 by the authors.ope

Vascular Regeneration With New Sources of Endothelial Cells

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Socio-Critical consciousness On Valerio Adami's Paintings

본 논문은 발레리오 아다미(Valerio Adami) 의 예술 세계에 나타난 사회비판의식에 관한 연구이다. 이탈리아 태생인 아다미는 특정 국가에 귀속되지 않는 예술가의 정체성을 바탕으로 인간과 사회의 관계를 모색하고, 이를 작품으로 형상화하였다. 본 논문은 현대인의 분열증적 소외에 대해 날카롭게 비판한 초기 작업 및 특정 인물들에 대한 공동체적 회고를 유도하는 후기 작업을 사회 비판적 메시지가 담긴 작품으로 분석하였으며, 더불어 인간 본연의 존엄성과 자유를 중요시하는 작가 자신의 사상을 예술 매체를 통해 관람자와 의사소통 하고자 한 점을 사회비판 의식의 표출로 주목하였다. 1960년대 작품들은 도시 사회의 익명적 공간이라고 할 수 있는 공공장소 속에 인간의 신체를 파편화 시키고, 성적인 욕망을 온전히 투여할 수 없는 부분적인 신체를 제시함으로써, 인간의 정체성을 박탈하고 성적 욕망을 억압하는 자본주의의 폭력성에 관해 서술한 것이다. 이 시기의 작품들은 아다미가 본격적인 작품 활동의 시작과 함께 몸을 담았던 신구상의 사회 비판적 입장을 공유하였음을 엿 볼 수 있는 것으로서, 이후 작품의 전개를 예비하는 초석이 되었다. 이시기 프랑스는 모든 가치체제의 변화를 시도하는 혼란과 반동의 시기로 규정 짓을 수 있다. 이러한 분위기는 예술분야에까지 그 파급 효과를 미치게 되었으며. 예술이 그 사회상을 반영한다는 측면에서 당연할 귀결로 이해할 수 있다. 특히 이 당시 미술의 사회적 기능으로서의 가치에 대한 중요성을 인식한 젊은 세대들은 예술전반에 대한 반성뿐만 아니라 사회 개혁의 의지를 미술로서 표현하고자 하였다. 이들은 당시의 정치적 이슈들을 작품의 주제로 삼아 공동작업을 하였을 뿐만 아니라, 자신들의 정치적 반향을 행동으로 실천함으로서 예술과 정치와의 접목을 지속적으로 추구하였다. 당시의 이러한 활동들은 '신구상(Nouvelle figuration)'이라는 용어로 지칭되어지며, 더불어 ' 서술적 구상' '비판적 구상' '정치적 구상' 등으로 일컬어졌는데, 이러한 다양한 명칭에서도 알 수 있듯이 프랑스의 예술의 흐름은 다양한 양상으로 전개되었음을 가늠하게 해준다. 특히 아다미는 1970년대에 자신만의 독특한 방법들을 제시하기 시작한다. 이 시기 작품들은 예술 매체를 비판 도구로 인식하는 초창기 태도에서 관람자와 작가자신 그리고 사회의 대립적인 관계에 대해 긍정적 방향으로 나아가기 위한 출구로 삼았다는 점에서 아다미의 보다 성숙한 예술관의 표출로 평가할 수 있다. 즉 그는 이시기에 초장기의 사회 비판적 태도에서 머무르지 않고 대립적인 인간 개체들의 화합과 화해의 장을 마련하고자 하였다. 아다미는 역사적 인물들과 특정 사건에 대한 객관적 자료 및 기록물에서 획득한 이미지에 작가 자신의 주관적 경험을 첨가하는 복합적 내러티브를 구축하였다. 이는 관람자의 정형화된 기억을 기억 메카니즘을 통해 자유롭게 확장시키는 주관적 내러티브 체계의 완성이라 할 수 있다. 객관적 자료와 함께 첨가된 상징적 모티브, 강렬한 원색. 변형된 문자 등의 조형적 요소들은 작가 자신의 사회비판 의식을 함축하고 있는 전달체인 동시에 관람자의 상상력을 자극하는 유도체이다. 따라서 이러한 불분명한 작가의 조형요소들은 관람자로 하여금 작품 해석에 있어서 능동적으로 역사적 인물과 사건에 대한 새로운 해석을 전개하도록 유도한다. 아다미 작품의 전반적인 특징이라 할 수 있는 형태의 단순성 및 해석의 모호함은 허구적 일상성을 조장하는 대중매체의 특정 형식에서 비롯되었다. 작가 자신의 메시지를 우회적으로 전달하기 위하여 고안해 낸 전략은 바로 만화적 패턴이었다. 즉 검은 윤곽선과 강렬한 원색을 통해 이루어지는 이미지의 불투명성과 직접성 초월이라는 아다미의 독특한 조형방식은 신구상 작가들의 직설적인 현실 비판적 작품들과 뚜렷이 구분되는 측면이라고 할 수 있다. 이러한 표현법에서도 특정한 국가, 사상, 예술사조에 종속되지 않으려는 작가의 보편적 정체성과 자유로운 탐구정신을 짐작할 수 있었다. 본 논문은 다양한 주제를 담고 있는 작품 전반을 인간과 사회 간 화해의 장으로 모색하기 위한 작가의 정치적 내러티브의 발전으로 분석하였다. 따라서 주제 속에 담겨진 사회비판 의식을 분석하고자 하는 본 논문은 아다미가 전달하고자 한 서술의 일관성을 분석하는 것에 그 목적이 있다.;This thesis focuses on socio-critical consciousness which was revealed in the narratives of Valerio Adami(1935 - )'s paintings. Adami, an artist born in Italy, concerned the relationship between modern men and society based on the free identity belongs to nowhere, and translate it into his works of art. This thesis analyzes varied themes in broad period works to lie the early one which criticizes nervous breakdown isolations in temporary men and to be the latter which induces the collective retrospect from certain well-known people involved in historical events. Moreover, the intention he conveyed human being's born nature dignity and freedom by meaningful works should be illuminated as a flux critical consciousness for society. Early works in 1960's depict the hidden violence which deprive of identities and suppress sexual desires in capitalism by dispersing a whole human body into segments in public space, anonymous places, such as a hotel, a restroom, and a pay phone booth, and exposing body parts that cannot contains eroticism. They show that Adami acquired critical sense, a fatal factor based on expectant works, from 'Nouvelle Figuratrion' that he participated in with starting his artistic career. The latter works in 1970's demonstrates the affluence and maturity of author's spirit. His early attitude was that he perceived a work of art as critical implement. However, he made his paintings an affirmative starting point later to soften the stiff relationship between an artist and spectators and other oppositive social ones. In other words, he moves towards reconciliation among respective individual conflicts. Adami constructed the complex narratives that were composed of images, acquired from documents and pictures on historical people and events, simultaneously , he added subjective experience to. This thesis illuminates his works of art to be the completion of subjective narratives that set one's fixed memories free. This procedure considerably resembles the flow of biological memorization mechanism. Symbolic motifs, appended to documents and pictures, intense primary colors, and artistic revised calligraphies are important elements that imply his critical sense and induce freely association of spectators. Therefore, every element of paintings in 1970's functions to help them construe voluntarily historical people and events. Simple shapes and ambiguous mood, inconsistent features of Adami's paintings, were originated from a certain mass media which exaggerates the falsehood of everyday life in capitalism. He invented his artistic strategy which is the rebirth of a cartoon pattern to transmit his critical message in a roundabout way. The opacity of images and transcendence of this world, accomplished by strong outlines and splendid primary colours, are feature differential from straightness, found in the other 'Nouvelle Figuratrion' painting. Author's universal identity and free spirit for pursuit are by no means subject to any boundary on nationalities, trends of thought, current artistic trends. This thesis analyze varied theme works as developing of author's critical narratives to produce the harmony between modern men and society. Therefore, this study aims at analysis the consistency of narratives that author has conveyed.목차 = ⅳ 논문개요 = ⅴ Ⅰ. 서론 = 1 Ⅱ. 신구상과 정치성 = 7 A. 1960년대 예술 - 정치 프로젝트 = 7 B. 아다미의 우회적 비판 = 12 Ⅲ. 비판적 서술에 따른 이미지 변형 = 17 A. 현대문명에 대한 비판 = 17 1. 인체의 파편화 = 19 2. 왜곡된 성 윤리에 대한 환유 = 27 B. 정치 이데올로기의 폭력성 고발 = 34 1. 시·공간의 초월적 구성 = 37 2. 사건 기록의 재해석 = 47 Ⅳ. 결론 = 58 참고문헌 = 62 도판목록 = 68 도판 = 75 Abstract = 8

랫트에서 새로운 발기부전 치료제 DA-8159와 nitroglycerin을 정맥주사 또는 경구투여하였을 때 약물역학적 상호작용 기전

Thesis(master`s)--서울대학교 대학원 :약학과 약물학전공,2005.Maste

Phospholipid-mediated Effects on Conformational Change and Fibrilllation of Amyloidogenic α-Synuclein

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Enhanced Therapeutic and Long-Term Dynamic Vascularization Effects of Human Pluripotent Stem Cell-Derived Endothelial Cells Encapsulated in a Nanomatrix Gel

BACKGROUND: Human pluripotent stem cell (hPSC)-derived endothelial cells (ECs) have limited clinical utility because of undefined components in the differentiation system and poor cell survival in vivo. Here, we aimed to develop a fully defined and clinically compatible system to differentiate hPSCs into ECs. Furthermore, we aimed to enhance cell survival, vessel formation, and therapeutic potential by encapsulating hPSC-ECs with a peptide amphiphile (PA) nanomatrix gel. METHODS: We induced differentiation of hPSCs into the mesodermal lineage by culturing on collagen-coated plates with a glycogen synthase kinase 3β inhibitor. Next, vascular endothelial growth factor, endothelial growth factor, and basic fibroblast growth factor were added for endothelial lineage differentiation, followed by sorting for CDH5 (VE-cadherin). We constructed an extracellular matrix-mimicking PA nanomatrix gel (PA-RGDS) by incorporating the cell adhesive ligand Arg-Gly-Asp-Ser (RGDS) and a matrix metalloproteinase-2-degradable sequence. We then evaluated whether the encapsulation of hPSC-CDH5+ cells in PA-RGDS could enhance long-term cell survival and vascular regenerative effects in a hind-limb ischemia model with laser Doppler perfusion imaging, bioluminescence imaging, real-time reverse transcription-polymerase chain reaction, and histological analysis. RESULTS: The resultant hPSC-derived CDH5+ cells (hPSC-ECs) showed highly enriched and genuine EC characteristics and proangiogenic activities. When injected into ischemic hind limbs, hPSC-ECs showed better perfusion recovery and higher vessel-forming capacity compared with media-, PA-RGDS-, or human umbilical vein EC-injected groups. However, the group receiving the PA-RGDS-encapsulated hPSC-ECs showed better perfusion recovery, more robust and longer cell survival (> 10 months), and higher and prolonged angiogenic and vascular incorporation capabilities than the bare hPSC-EC-injected group. Surprisingly, the engrafted hPSC-ECs demonstrated previously unknown sustained and dynamic vessel-forming behavior: initial perivascular concentration, a guiding role for new vessel formation, and progressive incorporation into the vessels over 10 months. CONCLUSIONS: We generated highly enriched hPSC-ECs via a clinically compatible system. Furthermore, this study demonstrated that a biocompatible PA-RGDS nanomatrix gel substantially improved long-term survival of hPSC-ECs in an ischemic environment and improved neovascularization effects of hPSC-ECs via prolonged and unique angiogenic and vessel-forming properties. This PA-RGDS-mediated transplantation of hPSC-ECs can serve as a novel platform for cell-based therapy and investigation of long-term behavior of hPSC-ECs.restrictio

Metal Ion Effect on the Conformational Change and Fibrillation of hIAPP Probed Using Electrospray Ionization Ion Mobility Mass Spectrometry

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Oligomeric States of hIAPP and the Effects on the Fibrillation in the Model Membrane System: Conformational Analysis using Mass Spectrometry and Spectroscopic Tools

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