Methods in molecular biology最新文献

Assessing Autophagy Flux in Glioblastoma Temozolomide Resistant Cells. 评估胶质母细胞瘤替莫唑胺耐药细胞的自噬通量

Methods in molecular biology Pub Date : 2024-09-28 DOI: 10.1007/7651_2024_571

Courtney Clark, Amir Barzegar Behrooz, Marco Cordani, Shahla Shojaei, Saeid Ghavami

引用次数: 0

Spontaneous Efficient Differentiation of Human Pluripotent Stem Cells (hPSC) Upon Co-culture of hPSCs with Human Neonatal Foreskin Fibroblasts in 3D. 人多能干细胞（hPSC）与人新生儿表皮成纤维细胞三维共培养后的自发高效分化。

Methods in molecular biology Pub Date : 2024-09-25 DOI: 10.1007/7651_2024_569

Muhammad Nihad, Sudheer Shenoy P, Bipasha Bose

{"title":"Spontaneous Efficient Differentiation of Human Pluripotent Stem Cells (hPSC) Upon Co-culture of hPSCs with Human Neonatal Foreskin Fibroblasts in 3D.","authors":"Muhammad Nihad, Sudheer Shenoy P, Bipasha Bose","doi":"10.1007/7651_2024_569","DOIUrl":"https://doi.org/10.1007/7651_2024_569","url":null,"abstract":"Pluripotent stem cells (PSCs) form well-formed embryoid bodies (EBs) in 3D culture. These EBs are formed in culture media lacking leukemia inhibitory factor (LIF) or basic fibroblast growth factor (bFGF) in mouse and human PSCs, respectively. EBs are excellent technical tools for understanding developmental biology and inducing controlled differentiation in succeeding experimental steps. Technically speaking, EBs are spontaneously differentiated PSCs in 3D and exhibit all three lineages in a time-point/sequential manner. For example, ectoderm will form first, followed by mesoderm and endoderm. We have attempted to co-culture human neonatal foreskin-derived fibroblast cells in our laboratory with the PSCs first in 2D conditions followed by the induction of EBs (PSC+fibroblasts co-cultured) in low attachment dishes. We also performed spontaneous differentiation of such EBs (co-cultured with fibroblasts). We checked the presence of markers of various lineages, namely, ectoderm, mesoderm, and endoderm in days 6, 10, and 12 day EBs. We have also compared the fibroblast co-cultured EBs, along with control EBs (derived from only PSCs). This co-culture system mimics the natural conditions of uterine implantation and the role of the endometrial fibroblasts in the induction of further embryonic development. The fibroblast co-cultured iPSC EBs had better roundness scores than the normal iPSC EBs and had a higher expression of lineage-specific markers.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142308110","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

Tubular Aggregates as a Marker of Aging in Skeletal Muscle. 作为骨骼肌衰老标志的管状聚集体

Methods in molecular biology Pub Date : 2024-09-25 DOI: 10.1007/7651_2024_567

Felipe Tadeu Galante Rocha de Vasconcelos, Brandow Willy Souza, Lucas Santos Souza, Mariz Vainzof

引用次数: 0

Polychromatic Flow Cytometry to Identify Rare Aged Hematopoietic Stem Cell Subpopulations. 多色流式细胞术识别罕见的老年造血干细胞亚群

Methods in molecular biology Pub Date : 2024-09-25 DOI: 10.1007/7651_2024_570

Natalia Skinder, Christina Pitsillidou, Alessandra Roberto, Gerald de Haan

引用次数: 0

Immunocompetent Brain Organoids with Microglia Allow Advanced Aging Research. 具有小胶质细胞的免疫功能脑有器官组织允许进行高级老化研究

Methods in molecular biology Pub Date : 2024-09-25 DOI: 10.1007/7651_2024_565

Raiane Oliveira Ferreira, Amanda Faria Assoni, Monize Valéria Ramos da Silva, Letícia Alves da Rocha, Mateus Vidigal de Castro, Débora Bertola, Mayana Zatz

引用次数: 0

Correction to: Monitoring Autophagy During Drosophila Oogenesis. 更正：果蝇卵子发生过程中的自噬监测

Methods in molecular biology Pub Date : 2024-09-21 DOI: 10.1007/7651_2024_572

Mrunmayee Kulkarni, Karan Selarka, Bhupendra V Shravage

引用次数: 0

Expansion and Maturation of Innate Lymphoid Cell Precursors Using Human iPSC-Derived Intestinal Organoids. 使用人类 iPSC 衍生的肠组织细胞扩增和成熟先天性淋巴细胞前体

Methods in molecular biology Pub Date : 2024-08-31 DOI: 10.1007/7651_2024_568

Emma Højmose Kromann, Geraldine M Jowett, Joana F Neves

{"title":"Expansion and Maturation of Innate Lymphoid Cell Precursors Using Human iPSC-Derived Intestinal Organoids.","authors":"Emma Højmose Kromann, Geraldine M Jowett, Joana F Neves","doi":"10.1007/7651_2024_568","DOIUrl":"https://doi.org/10.1007/7651_2024_568","url":null,"abstract":"Innate lymphoid cells (ILC) are enriched at mucosal barrier sites where they play critical roles in development and disease. Mucosal organoids offer a robust platform for the simultaneous differentiation and expansion of all subsets of mature ILC from a shared peripheral blood precursor. Critically, organoid identity drives tissue-specific imprinting of the culture-derived mature innate lymphoid cells, allowing for the study of bidirectional interactions between, e.g., intestinal organoids and intestine-specific ratios and populations of ILC. This protocol reduces the need for feeder cell lines and complex cytokine cocktails used to mature and maintain ILC, instead relying on a native niche of protein signals provided by mucosal epithelial cells. This protocol details the generation of human intestinal organoids (HIO) from human-induced pluripotent stem cells (hiPSC), and the subsequent establishment of co-cultures between HIO and ILC precursors for expansion and maturation. This approach has extensive applications for mechanistic studies of fundamental biological processes and as a potential GMP-compatible source of ILC for future cell therapies.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142109159","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

Quantifying Muscle Regeneration: Activated Muscle Satellite Cells and New Regenerated Myofibers in Chronic and Acute Degeneration Models. 量化肌肉再生：慢性和急性退化模型中的活化肌肉卫星细胞和新再生肌纤维。

Methods in molecular biology Pub Date : 2024-08-21 DOI: 10.1007/7651_2024_564

Antonio Fernando Ribeiro Junior, Brandow Willy Souza, Mariz Vainzof

{"title":"Quantifying Muscle Regeneration: Activated Muscle Satellite Cells and New Regenerated Myofibers in Chronic and Acute Degeneration Models.","authors":"Antonio Fernando Ribeiro Junior, Brandow Willy Souza, Mariz Vainzof","doi":"10.1007/7651_2024_564","DOIUrl":"https://doi.org/10.1007/7651_2024_564","url":null,"abstract":"Regeneration is a remarkable characteristic of the skeletal muscle. Triggered by common lesions, regeneration is stimulated resulting in muscle fiber repair and restoration of muscle homeostasis in normal muscle. In genetic dystrophic muscle, the cycle of degeneration/regeneration is an endless loop that leads to impaired regeneration and substitution of muscle fibers by connective and adipose tissue, causing muscle weakness. Identification and characterization of muscle regeneration steps can help discover potential therapy targets for muscle diseases and aging. Muscle regeneration markers such as the number of satellite cells in the muscle, the proportion of activated satellite cells, and the quantity of regenerating muscle fiber can be quantified using immunolabeling.Here we are presenting a quantitative method to measure muscle regeneration that can be applied to different proposals. To demonstrate the protocol applicability, we used models for acute and chronic muscle injuries. As model of acute degeneration, a wild-type C57BL6 mice with muscle injury induced by electroporation was used, and the muscle was analyzed after 5 and 10 days post-injury. DMDmdx mouse muscle was used as a model of chronic degeneration. The methodologies presented here are among the gold standard methodologies for muscle regeneration analysis and can be easily applied to any type of muscle regeneration study.","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142004591","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

Monitoring Autophagy During Drosophila Oogenesis. 监测果蝇卵子发生过程中的自噬作用

Methods in molecular biology Pub Date : 2024-08-10 DOI: 10.1007/7651_2024_563

Mrunmayee Kulkarni, Karan Selarka, Bhupendra V Shravage

引用次数: 0

The Chorioallantoic Membrane (CAM) Assay for the Analysis of Starvation-Induced Autophagy. 用于分析饥饿诱导的自噬的绒毛膜 (CAM) 分析法。

Methods in molecular biology Pub Date : 2024-08-10 DOI: 10.1007/7651_2024_562

Nazlı Şevval Menemenli, Özün Özcan, H Hazal Hüsnügil, Aliye Ezgi Güleç Taşkıran, Göksu Oral, Aytekin Akyol, Sreeparna Banerjee

引用次数: 0