Nascent matrix deposition supports alveolar organoid formation from aggregates in synthetic hydrogels DOI Creative Commons
Madeline K. Eiken, Charlie J. Childs,

Lindy K. Brastrom

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 19, 2024

Highlights Alveolar organoids are formed with a two-step, Matrigel-free method in semi-synthetic hyaluronic acid (HA) hydrogel The two-step offers control over alveolar size, density, and growth maintain their AT2 identity HA hydrogels secrete nascent extracellular matrix supporting organoid without Matrigel Summary Human induced pluripotent stem cell (iPSC) derived have emerged as system to model the epithelium homeostasis disease. However, typically grown Matrigel, mouse-sarcoma basement membrane that poor properties, prompting development of synthetic alternative. Here, we develop culture involves pre-aggregation hydrogel-based microwells followed by embedding supports growth, while also offering considerable properties. We find aggregated own (ECM) both upon hydrogels. Thus, gels described here allow us de-couple exogenous ECM order interrogate role formation.

Language: Английский

Biofunctionalized gelatin hydrogels support development and maturation of iPSC-derived cortical organoids DOI Creative Commons
Andrew Kjar,

Mia R Haschert,

José C. Zepeda

et al.

Cell Reports, Journal Year: 2024, Volume and Issue: 43(11), P. 114874 - 114874

Published: Oct. 17, 2024

Language: Английский

Citations

2
Biomaterial-assisted organoid technology for disease modeling and drug screening DOI Creative Commons

Yunyuan Shao,

Juncheng Wang,

Anqi Jin

et al.

Materials Today Bio, Journal Year: 2024, Volume and Issue: 30, P. 101438 - 101438

Published: Dec. 31, 2024

Language: Английский

Citations

2
A Hybrid 2D‐to‐3D in vitro Differentiation Platform Improves Outcomes of Cerebral Cortical Organoid Generation in hiPSCs DOI
Dosh Whye,

Erika M. Norabuena,

Gayathri Srinivasan

et al.

Current Protocols, Journal Year: 2024, Volume and Issue: 4(10)

Published: Oct. 1, 2024

Abstract Three‐dimensional (3D) cerebral cortical organoids are popular in vitro cellular model systems widely used to study human brain development and disease, compared traditional stem cell–derived methods that use two‐dimensional (2D) monolayer cultures. Despite the advancements made protocol for organoid derivation over past decade, limitations due biological, mechanistic, technical variables remain generating these complex 3D systems. Building from our previously established differentiation system, we have modifications existing resolve several of biological challenges when working with diverse groups induced pluripotent cell (hiPSC) lines. This improved blends a 2D culture format specification neural cells expansion neuroepithelial progenitor system self‐aggregation subsequent development. Furthermore, this “hybrid” approach is amenable both an accelerated as well alternative long‐term protocol. In addition establishing hybrid format, also offers phenotypic morphological characterization stage‐specific profiles using antibodies fluorescent‐based dyes live imaging. © 2024 Wiley Periodicals LLC. Basic Protocol 1 : hiPSC‐based into (NSCs) 2 Serial passaging (NPCs) Support Direct cryopreservation rapid thawing NSCs NPCs 3 Bulk aggregation neurospheres Alternate High‐throughput neurosphere formation migration assay LIVE/DEAD stain imaging 4 NeuroFluor NeuO dye

Language: Английский

Citations

2
Matrigel Tunes H9 Stem Cell-Derived Human Cerebral Organoid Development DOI Creative Commons
R. Chris Estridge,

Jennifer E. O’Neill,

Albert J. Keung

et al.

Organoids, Journal Year: 2023, Volume and Issue: 2(4), P. 165 - 176

Published: Oct. 5, 2023

Human cerebral organoids are readily generated from human embryonic stem cells and induced pluripotent useful in studying neurodevelopment. Recent work with have explored the creation of different brain regions impacts soluble mechanical cues. Matrigel is a gelatinous, heterogenous mixture extracellular matrix proteins, morphogens, growth factors secreted by Engelbreth-Holm-Swarm mouse sarcoma cells. It core component almost all organoid protocols, generally supporting neuroepithelial budding tissue polarization; yet, its roles effects beyond general requirement protocols not well understood, mode delivery variable, including embedding within it or form. Given widespread usage, we asked how H9 cell-derived hCO development composition affected dosage method. We found exposure influences size, morphology, cell type composition. also showed that greater amounts promote an increase number choroid plexus (ChP) cells, this regulated BMP4 pathway. These results illuminate on importance amount phenotype

Language: Английский

Citations

4
Nascent matrix deposition supports alveolar organoid formation from aggregates in synthetic hydrogels DOI Creative Commons
Madeline K. Eiken, Charlie J. Childs,

Lindy K. Brastrom

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 19, 2024

Highlights Alveolar organoids are formed with a two-step, Matrigel-free method in semi-synthetic hyaluronic acid (HA) hydrogel The two-step offers control over alveolar size, density, and growth maintain their AT2 identity HA hydrogels secrete nascent extracellular matrix supporting organoid without Matrigel Summary Human induced pluripotent stem cell (iPSC) derived have emerged as system to model the epithelium homeostasis disease. However, typically grown Matrigel, mouse-sarcoma basement membrane that poor properties, prompting development of synthetic alternative. Here, we develop culture involves pre-aggregation hydrogel-based microwells followed by embedding supports growth, while also offering considerable properties. We find aggregated own (ECM) both upon hydrogels. Thus, gels described here allow us de-couple exogenous ECM order interrogate role formation.

Language: Английский

Citations

1