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Guidelines for Manufacturing and Application of Organoids: Heart. 有机体的制造和应用指南:心脏
IF 2.5 4区 医学 Q3 CELL & TISSUE ENGINEERING Pub Date : 2024-05-30 Epub Date: 2024-05-23 DOI: 10.15283/ijsc24046
Hyang-Ae Lee, Dong-Hun Woo, Do-Sun Lim, Jisun Oh, C-Yoon Kim, Ok-Nam Bae, Sun-Ju Ahn

Cardiac organoids have emerged as invaluable tools for assessing the impact of diverse substances on heart function. This report introduces guidelines for general requirements for manufacturing cardiac organoids and conducting cardiac organoid-based assays, encompassing protocols, analytical methodologies, and ethical considerations. In the quest to employ recently developed three-dimensional cardiac organoid models as substitutes for animal testing, it becomes imperative to establish robust criteria for evaluating organoid quality and conducting toxicity assessments. This guideline addresses this need, catering to regulatory requirements, and describes common standards for organoid quality and toxicity assessment methodologies, commensurate with current technological capabilities. While acknowledging the dynamic nature of technological progress and the potential for future comparative studies, this guideline serves as a foundational framework. It offers a comprehensive approach to standardized cardiac organoid testing, ensuring scientific rigor, reproducibility, and ethical integrity in investigations of cardiotoxicity, particularly through the utilization of human pluripotent stem cell-derived cardiac organoids.

心脏器官模型已成为评估各种物质对心脏功能影响的宝贵工具。本报告介绍了制造心脏器官模型和进行基于心脏器官模型的检测的一般要求指南,包括操作规程、分析方法和伦理考虑因素。为了采用最新开发的三维心脏类器官模型来替代动物试验,当务之急是建立健全的类器官质量评估标准并进行毒性评估。本指南满足了这一需求,满足了监管要求,并描述了与当前技术能力相适应的类器官质量和毒性评估方法的通用标准。在承认技术进步的动态性和未来比较研究潜力的同时,本指南可作为一个基础框架。它为标准化心脏类器官测试提供了一种全面的方法,确保心脏毒性研究的科学严谨性、可重复性和道德完整性,特别是通过利用源自人类多能干细胞的心脏类器官。
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引用次数: 0
Guidelines for Packaging, Transport, and Storage of Source Cells for Organoids. 有机体源细胞的包装、运输和储存指南。
IF 2.3 4区 医学 Q3 CELL & TISSUE ENGINEERING Pub Date : 2024-05-30 Epub Date: 2024-05-13 DOI: 10.15283/ijsc24042
Sungin Lee, Dayeon Kwon, Han Byeol Lee, Sooyeon Jeon, Chihye Park, Tae Sung Kim, Jin Hee Lee, Il Ung Oh, Sun-Ju Ahn

This paper presents guidelines for the systematic management of packaging, storage, transportation, and traceability of source cells used for organoid research. Given the important role of source cells in organoid studies, it is important to ensure the preservation of their quality and integrity throughout transportation and distribution processes. The proposed guidelines, therefore, call for a cohesive strategy through these stages to minimize the risks of contamination, deterioration, and loss-threats that significantly compromise the safety, efficacy, and efficiency of source cells. Central to these guidelines is the quality control measures that include roles and responsibilities across the entire supply chain, with recommendations specific to packaging materials, transportation facilities, and storage management. Furthermore, the need for an integrated management system is emphasized, spanning from source cell collection to the final application. This system is crucial for maintaining the traceability and accountability of source cells, facilitating the sharing, distribution, and utilization on a global scale, and supporting to advance organoid research and development.

本文介绍了用于类器官研究的源细胞的包装、储存、运输和可追溯性的系统管理指南。鉴于源细胞在类器官研究中的重要作用,必须确保在整个运输和分销过程中保持其质量和完整性。因此,拟议指南要求在这些阶段采取协调一致的策略,最大限度地降低污染、变质和丢失的风险--这些威胁会严重影响源细胞的安全性、功效和效率。这些指南的核心是质量控制措施,其中包括整个供应链的角色和责任,以及针对包装材料、运输设施和储存管理的具体建议。此外,还强调了从源细胞采集到最终应用的综合管理系统的必要性。该系统对于保持源细胞的可追溯性和问责制,促进全球范围内的共享、分配和利用,以及支持推进类器官研究与开发至关重要。
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引用次数: 0
Guidelines for Manufacturing and Application of Organoids: Liver. 有机体制造和应用指南》:肝脏
IF 2.3 4区 医学 Q3 CELL & TISSUE ENGINEERING Pub Date : 2024-05-30 Epub Date: 2024-05-22 DOI: 10.15283/ijsc24044
Hye-Ran Moon, Seon Ju Mun, Tae Hun Kim, Hyemin Kim, Dukjin Kang, Suran Kim, Ji Hyun Shin, Dongho Choi, Sun-Ju Ahn, Myung Jin Son

Recent amendments to regulatory frameworks have placed a greater emphasis on the utilization of in vitro testing platforms for preclinical drug evaluations and toxicity assessments. This requires advanced tissue models capable of accurately replicating liver functions for drug efficacy and toxicity predictions. Liver organoids, derived from human cell sources, offer promise as a reliable platform for drug evaluation. However, there is a lack of standardized quality evaluation methods, which hinders their regulatory acceptance. This paper proposes comprehensive quality standards tailored for liver organoids, addressing cell source validation, organoid generation, and functional assessment. These guidelines aim to enhance reproducibility and accuracy in toxicity testing, thereby accelerating the adoption of organoids as a reliable alternative or complementary tool to animal testing in drug development. The quality standards include criteria for size, cellular composition, gene expression, and functional assays, thus ensuring a robust hepatotoxicity testing platform.

最近对监管框架的修订更加强调利用体外测试平台进行临床前药物评价和毒性评估。这就要求先进的组织模型能够准确复制肝脏功能,以预测药物疗效和毒性。肝脏器官组织来源于人体细胞,有望成为药物评估的可靠平台。然而,由于缺乏标准化的质量评价方法,阻碍了监管部门对其的认可。本文针对肝脏器官组织提出了全面的质量标准,涉及细胞源验证、器官组织生成和功能评估。这些指南旨在提高毒性测试的可重复性和准确性,从而加快采用器官组织作为药物开发中动物试验的可靠替代或补充工具。质量标准包括大小、细胞组成、基因表达和功能测试等方面的标准,从而确保建立一个稳健的肝毒性测试平台。
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引用次数: 0
Standards for Organoids. 有机体标准
IF 2.3 4区 医学 Q3 CELL & TISSUE ENGINEERING Pub Date : 2024-05-30 Epub Date: 2024-05-27 DOI: 10.15283/ijsc24043
Sun-Ju Ahn
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引用次数: 0
Guidelines for Manufacturing and Application of Organoids: Brain. 有机体制造和应用指南》:脑。
IF 2.3 4区 医学 Q3 CELL & TISSUE ENGINEERING Pub Date : 2024-05-30 Epub Date: 2024-05-23 DOI: 10.15283/ijsc24056
Taehwan Kwak, Si-Hyung Park, Siyoung Lee, Yujeong Shin, Ki-Jun Yoon, Seung-Woo Cho, Jong-Chan Park, Seung-Ho Yang, Heeyeong Cho, Heh-In Im, Sun-Ju Ahn, Woong Sun, Ji Hun Yang

This study offers a comprehensive overview of brain organoids for researchers. It combines expert opinions with technical summaries on organoid definitions, characteristics, culture methods, and quality control. This approach aims to enhance the utilization of brain organoids in research. Brain organoids, as three-dimensional human cell models mimicking the nervous system, hold immense promise for studying the human brain. They offer advantages over traditional methods, replicating anatomical structures, physiological features, and complex neuronal networks. Additionally, brain organoids can model nervous system development and interactions between cell types and the microenvironment. By providing a foundation for utilizing the most human-relevant tissue models, this work empowers researchers to overcome limitations of two-dimensional cultures and conduct advanced disease modeling research.

本研究为研究人员提供了有关脑器官组织的全面概述。它将专家意见与有关类器官定义、特征、培养方法和质量控制的技术摘要相结合。这种方法旨在提高脑器官组织在研究中的利用率。脑器官组织作为模拟神经系统的三维人类细胞模型,在研究人类大脑方面前景广阔。与传统方法相比,它具有复制解剖结构、生理特征和复杂神经元网络的优势。此外,脑组织器官还能模拟神经系统的发育以及细胞类型和微环境之间的相互作用。这项工作为利用与人类最相关的组织模型奠定了基础,使研究人员能够克服二维培养的局限性,开展先进的疾病建模研究。
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引用次数: 0
Gastric Organoid, a Promising Modeling for Gastric Stem Cell Homeostasis and Therapeutic Application. 胃有机体--胃干细胞稳态和治疗应用的前景模型
IF 2.3 4区 医学 Q3 CELL & TISSUE ENGINEERING Pub Date : 2024-05-03 DOI: 10.15283/ijsc23075
Subin Lee, Jang-Hyun Choi, So-Yeon Park, Jihoon Kim
The elucidation of the pathophysiology underlying various diseases necessitates the development of research platforms that faithfully mimic in vivo conditions. Traditional model systems such as two-dimensional cell cultures and animal models have proven inadequate in capturing the complexities of human disease modeling. However, recent strides in organoid culture systems have opened up new avenues for comprehending gastric stem cell homeostasis and associated diseases, notably gastric cancer. Given the significance of gastric cancer, a thorough understanding of its pathophysiology and molecular underpinnings is imperative. To this end, the utilization of patient-derived organoid libraries emerges as a remarkable platform, as it faithfully mirrors patient-specific characteristics, including mutation profiles and drug sensitivities. Furthermore, genetic manipulation of gastric organoids facilitates the exploration of molecular mechanisms underlying gastric cancer development. This review provides a comprehensive overview of recent advancements in various adult stem cell-derived gastric organoid models and their diverse applications.
要阐明各种疾病的病理生理学,就必须开发能忠实模拟体内条件的研究平台。事实证明,二维细胞培养和动物模型等传统模型系统不足以捕捉人类疾病模型的复杂性。然而,最近在类器官培养系统方面取得的进展为理解胃干细胞稳态和相关疾病(尤其是胃癌)开辟了新途径。鉴于胃癌的重要性,彻底了解其病理生理学和分子基础势在必行。为此,利用源自患者的类器官库成为一个重要的平台,因为它能忠实反映患者的特异性特征,包括突变特征和药物敏感性。此外,对胃癌类器官进行遗传操作有助于探索胃癌发展的分子机制。本综述全面概述了各种成体干细胞衍生胃有机体模型的最新进展及其多样化应用。
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引用次数: 0
Impaired Osteogenesis in Human Induced Pluripotent Stem Cells with Acetaldehyde Dehydrogenase 2 Mutations. 乙醛脱氢酶 2 基因突变的人类诱导多能干细胞成骨功能受损
IF 2.3 4区 医学 Q3 CELL & TISSUE ENGINEERING Pub Date : 2024-04-12 DOI: 10.15283/ijsc23151
Jooyoung Lim, Heeju Han, Se In Jung, Yeri Alice Rim, Ji Hyeon Ju
Acetaldehyde dehydrogenase 2 (ALDH2) is the second enzyme involved in the breakdown of acetaldehyde into acetic acid during the process of alcohol metabolism. Roughly 40% of East Asians carry one or two ALDH2*2 alleles, and the presence of ALDH2 genetic mutations in individuals may affect the bone remodeling cycle owing to accumulation of acetaldehyde in the body. In this study, we investigated the effects of ALDH2 mutations on bone remodeling. In this study, we examined the effects of ALDH2 polymorphisms on in vitro osteogensis using human induced pluripotent stem cells (hiPSCs). We differentiated wild-type (ALDH2*1/*1-) and ALDH2*1/*2-genotyped hiPSCs into osteoblasts (OBs) and confirmed their OB characteristics. Acetaldehyde was administered to confirm the impact caused by the mutation during OB differentiation. Calcium deposits formed during osteogenesis were significantly decreased in ALDH2*1/*2 OBs. The expression of osteogenic markers were also decreased in acetaldehyde-treated OBs differentiated from the ALDH2*1/*2 hiPSCs. Furthermore, the impact of ALDH2 polymorphism and acetaldehyde-induced stress on inflammatory factors such as 4-hydroxynonenal and tumor necrosis factor α was confirmed. Our findings suggest that individuals with ALDH2 deficiency may face challenges in acetaldehyde breakdown, rendering them susceptible to disturbances in normal bone remodeling therefore, caution should be exercised regarding alcohol consumption. In this proof-of-concept study, we were able to suggest these findings as a result of a disease-in-a-dish concept using hiPSCs derived from individuals bearing a certain mutation. This study also shows the potential of patient-derived hiPSCs for disease modeling with a specific condition.
乙醛脱氢酶 2(ALDH2)是酒精代谢过程中参与将乙醛分解为乙酸的第二种酶。约有 40% 的东亚人携带一个或两个 ALDH2*2 等位基因,个体中存在的 ALDH2 基因突变可能会因体内乙醛的积累而影响骨重塑周期。本研究调查了 ALDH2 基因突变对骨重塑的影响。在这项研究中,我们利用人体诱导多能干细胞(hiPSCs)研究了ALDH2多态性对体外成骨的影响。我们将野生型(ALDH2*1/*1-)和ALDH2*1/*2基因型的hiPSCs分化成成骨细胞(OBs),并确认了它们的OB特征。在 OB 分化过程中施用乙醛以确认突变造成的影响。在成骨过程中形成的钙沉积在ALDH2*1/*2 OBs中明显减少。在经乙醛处理的由ALDH2*1/*2 hiPSCs分化出的OB中,成骨标志物的表达也有所下降。此外,ALDH2多态性和乙醛诱导的应激对4-羟基壬烯醛和肿瘤坏死因子α等炎症因子的影响也得到了证实。我们的研究结果表明,ALDH2 缺乏症患者可能在乙醛分解方面面临挑战,使他们容易受到正常骨重塑的干扰,因此应谨慎饮酒。在这项概念验证研究中,我们利用从携带某种突变的个体中提取的 hiPSCs,通过 "皿中病 "概念得出了这些发现。这项研究还显示了患者来源的 hiPSCs 在特定疾病建模方面的潜力。
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引用次数: 0
Suppression of Glioblastoma Stem Cell Potency and Tumor Growth via LRRK2 Inhibition. 通过抑制 LRRK2 抑制胶质母细胞瘤干细胞的潜能和肿瘤生长
IF 2.3 4区 医学 Q3 CELL & TISSUE ENGINEERING Pub Date : 2024-04-08 DOI: 10.15283/ijsc24032
Saewhan Park, Kyung-Hee Kim, Yun-Hee Bae, Young Taek Oh, Hyemi Shin, Hyung Joon Kwon, Chan Il Kim, Sung Soo Kim, Hwan-Geun Choi, Jong Bae Park, Byoung Dae Lee
Leucine-rich repeat kinase 2 (LRRK2), a large GTP-regulated serine/threonine kinase, is well-known for its mutations causing late-onset Parkinson's disease. However, the role of LRRK2 in glioblastoma (GBM) carcinogenesis has not yet been fully elucidated. Here, we discovered that LRRK2 was overexpressed in 40% of GBM patients, according to tissue microarray analysis, and high LRRK2 expression correlated with poor prognosis in GBM patients. LRRK2 and stemness factors were highly expressed in various patient-derived GBM stem cells, which are responsible for GBM initiation. Canonical serum-induced differentiation decreased the expression of both LRRK2 and stemness factors. Given that LRRK2 is a key regulator of glioma stem cell (GSC) stemness, we developed DNK72, a novel LRRK2 kinase inhibitor that penetrates the blood-brain barrier. DNK72 binds to the phosphorylation sites of active LRRK2 and dramatically reduced cell proliferation and stemness factors expression in in vitro studies. Orthotopic patient-derived xenograft mouse models demonstrated that LRRK2 inhibition with DNK72 effectively reduced tumor growth and increased survival time. We propose that LRRK2 plays a significant role in regulating the stemness of GSCs and that suppression of LRRK2 kinase activity leads to reduced GBM malignancy and proliferation. In the near future, targeting LRRK2 in patients with high LRRK2-expressing GBM could offer a superior therapeutic strategy and potentially replace current clinical treatment methods.
富亮氨酸重复激酶 2 (LRRK2)是一种大型 GTP 调节丝氨酸/苏氨酸激酶,因其突变导致晚发性帕金森病而闻名。然而,LRRK2 在胶质母细胞瘤(GBM)癌变中的作用尚未完全阐明。在这里,我们根据组织芯片分析发现,40%的GBM患者体内LRRK2过表达,而LRRK2的高表达与GBM患者的不良预后相关。LRRK2和干性因子在各种来源于患者的GBM干细胞中高表达,这些干细胞是GBM发病的元凶。典型的血清诱导分化降低了LRRK2和干性因子的表达。鉴于LRRK2是胶质瘤干细胞(GSC)干性的关键调节因子,我们开发了一种新型LRRK2激酶抑制剂DNK72,它能穿透血脑屏障。DNK72能与活性LRRK2的磷酸化位点结合,在体外研究中能显著减少细胞增殖和干性因子的表达。患者异位移植小鼠模型表明,用 DNK72 抑制 LRRK2 能有效减少肿瘤生长并延长存活时间。我们认为,LRRK2 在调控 GSC 干性方面发挥着重要作用,抑制 LRRK2 激酶活性可降低 GBM 的恶性程度和增殖。在不久的将来,在高LRRK2表达的GBM患者中靶向LRRK2可提供一种更优越的治疗策略,并有可能取代目前的临床治疗方法。
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引用次数: 0
Crosstalk between Signaling Pathways and Energy Metabolism in Pluripotency. 多能性中信号通路与能量代谢之间的相互影响
IF 2.3 4区 医学 Q3 CELL & TISSUE ENGINEERING Pub Date : 2024-03-18 DOI: 10.15283/ijsc23173
Keun-Tae Kim, Seong-Min Kim, Hyuk-Jin Cha

The sequential change from totipotency to multipotency occurs during early mammalian embryo development. However, due to the lack of cellular models to recapitulate the distinct potency of stem cells at each stage, their molecular and cellular characteristics remain ambiguous. The establishment of isogenic naïve and primed pluripotent stem cells to represent the pluripotency in the inner cell mass of the pre-implantation blastocyst and in the epiblast from the post-implantation embryo allows the understanding of the distinctive characteristics of two different states of pluripotent stem cells. This review discusses the prominent disparities between naïve and primed pluripotency, including signaling pathways, metabolism, and epigenetic status, ultimately facilitating a comprehensive understanding of their significance during early mammalian embryonic development.

从全能性到多能性的顺序变化发生在哺乳动物胚胎的早期发育过程中。然而,由于缺乏细胞模型来再现干细胞在每个阶段的不同潜能,它们的分子和细胞特征仍然模糊不清。建立同源的幼稚多能干细胞和原始多能干细胞,代表植入前胚泡内细胞团和植入后胚胎上胚层的多能性,有助于了解两种不同状态的多能干细胞的独特特征。这篇综述讨论了幼稚多能性和原始多能性之间的显著差异,包括信号通路、新陈代谢和表观遗传状态,最终有助于全面了解它们在哺乳动物早期胚胎发育过程中的意义。
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引用次数: 0
Stem Cell-Based Approaches in Parkinson's Disease Research. 基于干细胞的帕金森病研究方法。
IF 2.3 4区 医学 Q3 CELL & TISSUE ENGINEERING Pub Date : 2024-03-07 DOI: 10.15283/ijsc23169
Min Seong Kim, Subeen Yoon, Jiwoo Choi, Yong Jun Kim, Gabsang Lee

Parkinson's disease (PD) is a neurodegenerative condition characterized by the loss of midbrain dopaminergic neurons, leading to motor symptoms. While current treatments provide limited relief, they don't alter disease progression. Stem cell technology, involving patient-specific stem cell-derived neurons, offers a promising avenue for research and personalized regenerative therapies. This article reviews the potential of stem cell-based research in PD, summarizing ongoing efforts, their limitations, and introducing innovative research models. The integration of stem cell technology and advanced models promises to enhance our understanding and treatment strategies for PD.

帕金森病(PD)是一种神经退行性疾病,其特征是中脑多巴胺能神经元的丧失,从而导致运动症状。虽然目前的治疗方法能提供有限的缓解,但无法改变疾病的进展。干细胞技术涉及患者特异性干细胞衍生神经元,为研究和个性化再生疗法提供了一条前景广阔的途径。本文回顾了基于干细胞的帕金森病研究潜力,总结了正在进行的工作及其局限性,并介绍了创新研究模型。干细胞技术与先进模型的结合有望增强我们对帕金森病的理解和治疗策略。
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引用次数: 0
期刊
International journal of stem cells
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