Methods in molecular biology最新文献

Patient-Derived Bladder Tumor Organoids Isolation and Culture: Conventional and Cost-Reduction Strategy. 患者来源的膀胱肿瘤类器官分离和培养：传统和降低成本的策略。

Methods in molecular biology Pub Date : 2025-09-13 DOI: 10.1007/7651_2025_664

Mahsa Mollapour Sisakht, Shirin Hekmatirad

{"title":"Patient-Derived Bladder Tumor Organoids Isolation and Culture: Conventional and Cost-Reduction Strategy.","authors":"Mahsa Mollapour Sisakht, Shirin Hekmatirad","doi":"10.1007/7651_2025_664","DOIUrl":"https://doi.org/10.1007/7651_2025_664","url":null,"abstract":"Organoids are three-dimensional structures generated in vitro from tissue samples, induced pluripotent stem cells (iPSCs), and/or adult stem cells. Patient-derived organoids (PDOs) represent one of the most physiologically relevant culture systems, closely recapitulating the histological and functional features of the original tissue. They can be established in the laboratory for various applications, including regenerative medicine, drug screening, personalized medicine, and targeted therapy. Since 2009, organoid isolation and culture protocols have been reported for multiple organs, such as the colon, stomach, liver, lung, brain, breast, and bladder. Here, we describe a protocol for the isolation and culture of bladder tumor organoids derived from patients undergoing cystectomy or transurethral resection of bladder tumor (TUR). In addition to the conventional methodology, we introduce a cost-effective alternative approach utilizing sodium alginate hydrogel and fibroblast-conditioned medium (FCM). This strategy offers a reproducible, xeno-free, and low-cost platform that is well suited for both clinical research and resource-limited laboratory settings.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145054770","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Live Cell Imaging for Keratinocyte Lineage Tracing. 角质形成细胞谱系追踪的活细胞成像。

Methods in molecular biology Pub Date : 2025-08-06 DOI: 10.1007/7651_2025_656

Brook Abegaze, Brooke Vittimberga, Nwamaka Ijeh, Giselle Vitcov, Richard Kim, Ruby Ghadially

引用次数: 0

Extracellular Vesicle Production from Human Blood Vessel Organoids in a Vertical Wheel Bioreactor. 在立式轮式生物反应器中制备人血管类器官的细胞外囊泡。

Methods in molecular biology Pub Date : 2025-08-05 DOI: 10.1007/7651_2025_661

Justice Ene, Falak Syed, Shaoyang Ma, Shaoxuan Ma, Sailesti Joshi, Yan Li

{"title":"Extracellular Vesicle Production from Human Blood Vessel Organoids in a Vertical Wheel Bioreactor.","authors":"Justice Ene, Falak Syed, Shaoyang Ma, Shaoxuan Ma, Sailesti Joshi, Yan Li","doi":"10.1007/7651_2025_661","DOIUrl":"https://doi.org/10.1007/7651_2025_661","url":null,"abstract":"Although distinguished for their differentiation capacity, human-induced pluripotent stem cells (iPSCs)-derived extracellular vesicles (EVs) have been shown to contribute to functional recovery in the treatment of various traumatic and degenerative diseases. This promising role in therapeutic applications has resulted in considerable attention aimed toward their effective bio-manufacturing. However, traditional culture systems face various insufficiencies. Planar 2D culture results in a lack of scalability, with difficulty in manufacturing clinically relevant doses. Additionally, planar 2D culture lacks the complexity of in vivo biological systems. Although organoids have been proposed to fit this gap by better mimicking in vivo conditions, the traditional generation method of using static culture results in inefficient nutrient and waste transfer. Earlier bioreactor systems, which aim to resolve these issues, also face limitations of homogeneity and stress. Thus, vertical wheel bioreactors (VWBRs) with low shear stress profiles have recently emerged for stem cell organoid cultures, resulting in a more efficient and true-to-form manufacturing process for the secreted EVs. In this chapter, we describe an approach to generate and quantify EVs secreted by iPSC-differentiated human blood vessel organoids (iBVOs) grown in a scalable VWBR. iPSCs are expanded and then differentiated into iBVOs with differentiation media in VWBRs. Their produced EVs are subsequently isolated from the media and quantified using nanoparticle tracking analysis. This culture system should be able to produce a large quantity of the iBVO-derived EVs for the subsequent preclinical and clinical applications.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144784647","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Derivation of Lung Organoids from Human-Induced Pluripotent Stem Cells for Respiratory Infection Studies. 人诱导多能干细胞衍生肺类器官用于呼吸道感染研究。

Methods in molecular biology Pub Date : 2025-07-31 DOI: 10.1007/7651_2025_660

Sandra L Leibel

引用次数: 0

Studying Hair Growth in Mice: Synchronization of Hair Follicle Growth by Depilation. 小鼠毛发生长研究：脱毛对毛囊生长的同步作用。

Methods in molecular biology Pub Date : 2025-07-31 DOI: 10.1007/7651_2025_653

Wei-Hung Wang, Renzhi Hou, Tyng-Shiuan Hsieh, Maksim V Plikus, Sung-Jan Lin

{"title":"Studying Hair Growth in Mice: Synchronization of Hair Follicle Growth by Depilation.","authors":"Wei-Hung Wang, Renzhi Hou, Tyng-Shiuan Hsieh, Maksim V Plikus, Sung-Jan Lin","doi":"10.1007/7651_2025_653","DOIUrl":"https://doi.org/10.1007/7651_2025_653","url":null,"abstract":"Hair follicles manifest distinct morphological, cellular, and molecular features as they progress through active growth (anagen), regression (catagen), and rest (telogen) phases of regenerative cycles. Since hair growth stalls in vitro and because numerous skin-specific murine genetic tools are readily available, studies on hair growth are commonly performed in mice in vivo. In such murine studies, it is often necessary to determine accurate hair cycle stages and to obtain large numbers of synchronized hair follicles at predefined experimental time points. These goals are hindered by the fact that natural hair growth in mice is temporally and spatially asynchronous. Thus, artificial hair growth synchronization by means of easy-to-perform hair depilation is a commonly used technique. Hair depilation rapidly resets hair cycle, such that skin with uniform anagen, catagen, or telogen hair follicles can be reliably collected from mice at specific post-depilation experimental time points. Further, progression of hair growth cycle after depilation can be monitored non-invasively in mice and compared between mutant and control mice. This is achieved through observing and recording hair pigmentation-driven changes in skin color tone. In this chapter, we discuss technical aspects of performing hair depilation procedure, commonly used experimental means for post-depilation hair growth analyses, as well as the limitations of the depilation method.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144742602","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Isolation, Transplantation, and Long-Term Noninvasive Tracking of DiD-Labeled EpCAM+ Human Fetal Hepatic Progenitors in Mouse Livers. 小鼠肝脏中did标记EpCAM+人胎肝祖细胞的分离、移植和长期无创追踪。

Methods in molecular biology Pub Date : 2025-07-22 DOI: 10.1007/7651_2025_658

Chaturvedula Tripura, Sandeep Kumar Vishwakarma, Srinivas Gunda

{"title":"Isolation, Transplantation, and Long-Term Noninvasive Tracking of DiD-Labeled EpCAM+ Human Fetal Hepatic Progenitors in Mouse Livers.","authors":"Chaturvedula Tripura, Sandeep Kumar Vishwakarma, Srinivas Gunda","doi":"10.1007/7651_2025_658","DOIUrl":"https://doi.org/10.1007/7651_2025_658","url":null,"abstract":"Chronic liver disease (CLD) is a progressive condition characterized by the deterioration of liver structure and function, resulting from persistent injury and inflammation. Liver cell therapy has emerged as a promising alternative bridging strategy for patients waiting for the availability of a suitable donor liver for transplantation. Fetal human hepatic progenitor cells (fHPCs) hold great potential as a source for liver regeneration and restoration of liver function in individuals with CLD. A key challenge in liver cell therapy lies in the ability to effectively track transplanted donor cells, monitoring their homing, repopulation, and functional integration into the recipient's liver.This protocol outlines a comprehensive methodology for isolating fHPCs, enrichment of EpCAM positive cells, and labeling them with DiD dye. It also details the procedure for inducing liver fibrosis in SCID mice, transplanting the donor fHPCs, and conducting noninvasive, long-term imaging to track the transplanted cells in recipient SCID mice. Furthermore, we outline a thorough approach to confirm the presence and functional integration of the transplanted cells within the recipient livers.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144675259","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A Deep Learning Approach to Assessing Cell Identity in Stem Cell-Based Embryo Models. 基于干细胞的胚胎模型中评估细胞身份的深度学习方法。

Methods in molecular biology Pub Date : 2025-07-22 DOI: 10.1007/7651_2025_654

Nazmus Salehin, Martin Proks, Joshua M Brickman

引用次数: 0

Electrospun Poly(vinyl alcohol)/Silver Nitrate (PVA/AgNO₃) Nanofibers Incorporated with Mesenchymal Stem Cells for Wound Dressing Applications. 电纺丝聚（乙烯醇）/硝酸银（PVA/AgNO₃）纳米纤维与间充质干细胞结合用于伤口敷料应用。

Methods in molecular biology Pub Date : 2025-07-17 DOI: 10.1007/7651_2025_659

Aysegul Tiryaki, Ayse Ceren Calikoglu-Koyuncu

引用次数: 0

Generation of the Nonintegrated Human Bilaminar Embryo Model (Bilaminoid) from Naïve Pluripotent Stem Cells. 从Naïve多能干细胞生成非整合人双层胚胎模型（Bilaminoid）。

Methods in molecular biology Pub Date : 2025-07-15 DOI: 10.1007/7651_2025_652

Takumi Okubo, Yasuhiro Takashima