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2.5D tubulogenesis

ABSTRACT: Hepatocyte growth factor-induced three-dimensional tubulogenesis (3D) is a simple and highly controllable system for studying epithelial tubule initiation and maintenance. However, due to its limited efficiency and asynchronous development, isolating genes associated with specific morphological changes observed during tubule formation has been unfeasible. Here we report a significantly enhanced in vitro culture method called 2.5-dimensional tubulogenesis (2.5D). Detailed image analysis of 2.5D has revealed morphologically distinct stages -- monolayer, extension, and tubule -- and finds that these stages display in a highly synchronized manner. Using time-course transcriptional profiling, we collected genes whose expression changes specifically associate with extension and tubule stage.

ORGANISM(S): Canis lupus familiaris

PROVIDER: GSE25514 | GEO | 2010/12/24

SECONDARY ACCESSION(S): PRJNA133765

REPOSITORIES: GEO

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2.5D tubulogenesis

Project description:Hepatocyte growth factor-induced three-dimensional tubulogenesis (3D) is a simple and highly controllable system for studying epithelial tubule initiation and maintenance. However, due to its limited efficiency and asynchronous development, isolating genes associated with specific morphological changes observed during tubule formation has been unfeasible. Here we report a significantly enhanced in vitro culture method called 2.5-dimensional tubulogenesis (2.5D). Detailed image analysis of 2.5D has revealed morphologically distinct stages -- monolayer, extension, and tubule -- and finds that these stages display in a highly synchronized manner. Using time-course transcriptional profiling, we collected genes whose expression changes specifically associate with extension and tubule stage. Time-course microarrays to identify genes whose temporal regulation is associated with extension and tubule stages. Four biological replicates for each stage (monolayer, extension, and tubule).

2010-12-24 | E-GEOD-25514 | biostudies-arrayexpress

Canis lupus familiaris

Project description:2.5D tubulogenesis

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2D and 3D Madin-Darby Canine Kidney Cell Type II cell line response to hepatocyte growth factor

Project description:Analysis of 2D (transwell) and 3D (collagen type I) cultured MDCK cells and HGF (a MAPK activator). Traditional 2D cultures are fast and inexpensive but do no mimic natural niche/cell environment as well as the more laborious and costly 3D-cultures. 3D cultures, arguably, are better models for the study of developmental processes, such as tubulogenesis. Epithelial organs (such as kidney) develop via tubulogenesis, a process, at least in part, regulated by MAPK signaling. Therefore, 2D and 3D cells also treated with HGF plus MAPK inhibitors. Results provide insights into differential response to HGF-induced tubulogenesis depending on cell culture conditions (2D vs. 3D).

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Project description:Purpose: The goals of this study are to define transcriptome (RNA-seq) of mouse preimplantation embryos at different stages of development under a range of different environmental conditions. Methods: Mouse preimplantation embro transcriptional profiles were generated using embryos at several different developmental stages using Smart-seq2. Results: RNA-seq analysis finds that there is a highly dynamic pattern of gene expression during the preimplantation period. The sensitiivty to nutrient conditions varies markedly at different stages of development, with 2C embryos more sensitive to pyruvate omission than later stage embryos. Conclusions: Our study represents a comprehensive analysis of the mouse preimplantation development transcriptome, and how pyruvate provision impacts different developmental stages.

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2D and 3D Madin-Darby Canine Kidney Cell Type II cell line response to hepatocyte growth factor

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