MAGIC matrices: freeform bioprinting materials to support complex and reproducible organoid morphogenesis DOI Creative Commons
Austin J. Graham, Michelle W.L. Khoo, Vasudha Srivastava

et al.

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

Published: Feb. 5, 2024

Organoids are powerful models of tissue physiology, yet their applications remain limited due to relatively simple morphology and high organoid-to-organoid structural variability. To address these limitations we developed a soft, composite yield-stress extracellular matrix that supports optimal organoid morphogenesis following freeform 3D bioprinting cell slurries at tissue-like densities. The material is designed with two temperature regimes: 4 °C it exhibits reversible behavior support long printing times without compromising viability. When transferred culture 37 °C, the cross-links similar viscoelasticity plasticity basement membrane extracts such as Matrigel. We first characterize rheological properties MAGIC matrices optimize morphogenesis, including low stiffness stress relaxation. Next, combine this custom piezoelectric printhead allows more reproducible robust self-organization from uniform spatially organized "seeds." apply for high-throughput generation intestinal, mammary, vascular, salivary gland, brain arrays structurally those grown in pure Matrigel, but exhibit dramatically improved homogeneity size, shape, maturation time, efficiency morphogenesis. flexibility method enabled fabrication fully microphysiological systems, perfusable tubes experience cyclic strain response pressurization. Furthermore, reproducibility structure increased statistical power drug assay by up 8 orders-of-magnitude given number comparisons. Combined, advances lay foundation efficient complex morphologies canalizing both space time.

Language: Английский

Guidelines for Manufacturing and Application of Organoids: Liver DOI Open Access
Hye‐ran Moon, Seon Ju Mun, Tae Hun Kim

et al.

International Journal of Stem Cells, Journal Year: 2024, Volume and Issue: 17(2), P. 120 - 129

Published: May 22, 2024

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 accurately replicating liver functions efficacy predictions. Liver organoids, derived from human cell sources, offer promise as reliable platform evaluation. However, there is lack standardized quality evaluation methods, which hinders their acceptance. paper proposes comprehensive standards tailored addressing source validation, organoid generation, functional assessment. These guidelines aim enhance reproducibility accuracy testing, thereby accelerating adoption organoids alternative or complementary tool animal development. The include criteria size, cellular composition, gene expression, assays, thus ensuring robust hepatotoxicity platform.

Language: Английский

Citations

4
Pheno‐Morphological Screening and Acoustic Sorting of 3D Multicellular Aggregates Using Drop Millifluidics DOI Creative Commons

Leon Rembotte,

Thomas Beneyton, Lionel Buisson

et al.

Advanced Science, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 10, 2025

Abstract Three‐dimensional multicellular aggregates (MCAs) like organoids and spheroids have become essential tools to study the biological mechanisms involved in progression of diseases. In cancer research, they are now widely used as vitro models for drug testing. However, their analysis still relies on tedious manual procedures, which hinders routine use large‐scale assays. Here, a novel drop millifluidic approach is introduced screen sort large populations containing over one thousand MCAs: ImOCAS (Image‐based Organoid Cytometry Acoustic Sorting). This system utilizes real‐time image processing detect pheno‐morphological traits MCAs. They then encapsulated millimetric drops, actuated on‐demand using acoustic radiation force. The performance demonstrated by sorting with uniform sizes from heterogeneous population, isolating different phenotypes. lays groundwork high‐throughput screening high‐content MCAs controlled morphological phenotypical properties, promises accelerated progress biomedical research.

Language: Английский

Citations

0
Advances in lacrimal gland organoid development: Techniques and therapeutic applications DOI
Ahmed Abdal Dayem, Soo Bin Jang,

Nahee Lim

et al.

Biomedicine & Pharmacotherapy, Journal Year: 2025, Volume and Issue: 183, P. 117870 - 117870

Published: Jan. 26, 2025

Language: Английский

Citations

0
An in vitro model for cardiac organoid production: the combined role of geometrical confinement and substrate stiffness. DOI Creative Commons
Rosaria Santoro, Luca Piacentini, Chiara Vavassori

et al.

Materials Today Bio, Journal Year: 2025, Volume and Issue: 31, P. 101566 - 101566

Published: Feb. 15, 2025

Induced pluripotent stem cells (iPSCs), carrying the patient's genetic background, open path to advanced in vitro modeling. The feasibility of recapitulating complex pathophysiological scenarios depends on iPSC's ability differentiate into plurality specific organ resident cells, their maturation and networking. To this end, a strong interest has arisen organoids, 3D structures, obtained by exploiting iPSC natural capability self-assemble rebuild parts. In study, we describe characterization novel iPSC-based cardiac organoid (CO) model, generated high-throughput cost-effective method. Organoids were culture onto substrates known stiffness, under geometrical confinement inducing differentiation small-molecule-based modulation Wnt pathway. COs characterized using multi-omic approach (including bulk/single-cell RNA-sequencing, proteomic analysis), immunofluorescence, electrophysiology (patch clamp), optical recording-based contraction measurements. Results showed that recapitulate relevant features, including spontaneous contraction, multicellularity (e.g., cardiomyocytes, fibroblasts, epicardial layer) chamber organization. Moreover, environmental mechanical cues significant effect features. particular, culturing organoids low range characterizing embryonal surrounding, enriched gene sets related maturity cardiomyocyte ultrastructure. Functionally, different cardiac-specific ionic currents consistent action potentials recorded upon patch-clamp cardiomyocytes dissociated from COs. Finally, beating rate whole was monitored non-destructively via video recording quantified, demonstrating response clinically used chronotropic compounds, supporting future implementation proposed as platform for drug testing.

Language: Английский

Citations

0
A perfused iPSC-derived proximal tubule model for predicting drug-induced kidney injury DOI Creative Commons
Matthias Lechtenberg, Coraline Chéneau, Kevin Riquin

et al.

Toxicology in Vitro, Journal Year: 2025, Volume and Issue: unknown, P. 106038 - 106038

Published: Feb. 1, 2025

Language: Английский

Citations

0
A novel approach for lymphatic organoid embedding: eosin pre-staining and agarose pre-embedding DOI Creative Commons

Bao-Feng Wang,

Yingying Wang,

Yun-Lan Yi

et al.

Tissue Barriers, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 26, 2025

Adenoid organoids, as the primary immune barrier of airway, provide valuable models for studying lymphatic tissue function, but their histological processing remains challenging due to fragile structure and lack adhesion. Here, we introduce a novel approach that combines eosin pre-staining with agarose pre-embedding enhance visibility structural integrity during paraffin embedding. This method simplifies sectioning improves quality hematoxylin (HE) immunofluorescence (IF) staining, yielding clear stable signals. By addressing key limitations in organoid processing, this technique provides reliable solution IF studies, facilitating future research on adenoid organoids.

Language: Английский

Citations

0
Longitudinal and large-scale monitoring of transcriptome and RBP-RNA interactome in living cells by engineered protein nanocages DOI Creative Commons

Yangming Wang,

Lu‐Feng Hu,

Gang Xie

et al.

Research Square (Research Square), Journal Year: 2025, Volume and Issue: unknown

Published: March 26, 2025

Abstract Nondestructive sequencing of RNA from live cells is essential for monitoring and understanding dynamic biological processes. However, most existing methods rely on cell lysis or fixation, limiting their applicability longitudinal studies. Here, we introduce POND-seq (Protein nanocage-empOwered Non-Destructive sequencing), a novel approach that employs secretory protein nanocages fused with RNA-binding proteins (RBPs) to capture the RBP-RNA interactome transcriptome in cells. reliably identifies targets canonical RBPs across multiple types. By fusing poly(A)-binding (PABPC1) nanocage, demonstrate can monitor transcriptomic changes response signaling stimuli selectively cell-type-specific transcriptomes mixed populations. Additionally, facilitates dissection domains key amino acid residues critical interactions. We further highlight its utility large-scale screening, offering compelling evidence pathogenicity FMR1 variants. represents transformative advancement biology, biology precision medicine, enabling unprecedented insights into cellular dynamics disease mechanisms.

Language: Английский

Citations

0
Robust and reproducible human intestinal organoid-derived monolayer model for analyzing drug absorption DOI Creative Commons
Kai Tanaka,

Tatsuki Mochizuki,

Shogo Baba

et al.

Scientific Reports, Journal Year: 2025, Volume and Issue: 15(1)

Published: April 3, 2025

Predicting the absorption of orally administered drugs is crucial to drug development. Current in vitro models lack physiological relevance, robustness, and reproducibility, thus hindering reliable predictions. In this study, we developed a reproducible robust culture method generate human intestinal organoid-derived monolayer model that can be applied study through step-by-step approach. Our showed similarity primary enterocytes terms absorption-related gene expression profile, tight barrier function, tolerability toward artificial bile juice, transporter metabolizing enzyme nuclear receptor activity. This organoids derived from multiple donors. The permeability launched 19 our demonstrated correlation with Fa values, an R2 value 0.88. Additionally, by combining modeling simulation approaches, estimated FaFg values for seven out nine drugs, including CYP3A substrates, fell within 1.5 times range values. Applying discovery process might bridge gap between preclinical clinical research increase success rates

Language: Английский

Citations

0
Organoids and organs-on-chips: Recent advances, applications in drug development, and regulatory challenges DOI
Liangbin Zhou, Jingjing Huang, Cun Li

et al.

Med, Journal Year: 2025, Volume and Issue: 6(4), P. 100667 - 100667

Published: April 1, 2025

Language: Английский

Citations

0
Reproducibility of PD patient-specific midbrain organoid data for in vitro disease modelling DOI Creative Commons
Elisa Zuccoli,

Haya Al Sawaf,

Mona Tuzza

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: April 21, 2025

Abstract Midbrain organoids are advanced in vitro cellular models for disease modelling. They have been used successfully over the past decade Parkinson’s (PD) research and drug development. The three-dimensional structure multicellular composition allow under more physiological conditions than is possible with conventional 2D models. However, there concerns field regarding organoid batch-to-batch variability thus reproducibility of results. In this manuscript, we generate multiple independent midbrain batches derived from healthy individuals or GBA-N370S mutation-carrying PD patients to evaluate mutation-associated transcriptomic metabolic signature as well selected protein abundance. Our analysis shows that GBA-PD-associated phenotypes reproducible across generation time points. This proves not only suitable modelling, but also represent robust highly

Language: Английский

Citations

0