Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Rapid Assessment of Italian Honey Chemical Composition and Botanical Origin Using NIR Spectroscopy Coupled with Chemometric Analysis.

Sensors (Basel, Switzerland)·2026
Same author

Xanthan Gum-Iron System: Natural, Mechanically Tunable, Bioactive, and Magnetic-Responsive Hydrogels for Biomedical Engineering Applications.

ACS applied materials & interfaces·2025
Same author

Electrospun Polymer Fiber Mats for Persulfide Prodrug Delivery.

Biomacromolecules·2025
Same author

A Proteomic Approach to Determine Stem Cell Skeletal Differentiation Signature on Additive Manufactured Scaffolds.

Small science·2025
Same author

3D Niche-Inspired Scaffolds as a Stem Cell Delivery System for the Regeneration of the Osteochondral Interface.

Advanced materials (Deerfield Beach, Fla.)·2024
Same author

An <i>in vitro</i> model system based on calcium- and phosphate ion-induced hMSC spheroid mineralization.

Materials today. Bio·2023

Related Experiment Video

Updated: Mar 17, 2026

Author Spotlight: Advancements in Stem Cell Regenerative Therapy Through Photobiomodulation
05:42

Author Spotlight: Advancements in Stem Cell Regenerative Therapy Through Photobiomodulation

Published on: April 5, 2024

1.6K

3D Soft Hydrogels Induce Human Mesenchymal Stem Cells "Deep" Quiescence.

David Boaventura Gomes1, Timo Rademakers1, Ana Filipa Henriques Lourenço1

  • 1Complex Tissue Regeneration Department, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Maastricht, the Netherlands.

Advanced Healthcare Materials
|March 16, 2026
PubMed
Summary

Developing a 3D alginate hydrogel with RGD improved human mesenchymal stem cells (hMSCs) phenotype. These 3D cultures induced deep quiescence, unlike 2D cultures, offering insights for cell therapies.

Keywords:
3D cultureG0RAPTORadult stem cellsdeep quiescencemTORmesenchymal stem cellmesenchymal stromal cellquiescence

More Related Videos

Cellular Encapsulation in 3D Hydrogels for Tissue Engineering
09:37

Cellular Encapsulation in 3D Hydrogels for Tissue Engineering

Published on: October 26, 2009

37.7K
Preparation of Chitosan-based Injectable Hydrogels and Its Application in 3D Cell Culture
08:05

Preparation of Chitosan-based Injectable Hydrogels and Its Application in 3D Cell Culture

Published on: September 29, 2017

20.2K

Related Experiment Videos

Last Updated: Mar 17, 2026

Author Spotlight: Advancements in Stem Cell Regenerative Therapy Through Photobiomodulation
05:42

Author Spotlight: Advancements in Stem Cell Regenerative Therapy Through Photobiomodulation

Published on: April 5, 2024

1.6K
Cellular Encapsulation in 3D Hydrogels for Tissue Engineering
09:37

Cellular Encapsulation in 3D Hydrogels for Tissue Engineering

Published on: October 26, 2009

37.7K
Preparation of Chitosan-based Injectable Hydrogels and Its Application in 3D Cell Culture
08:05

Preparation of Chitosan-based Injectable Hydrogels and Its Application in 3D Cell Culture

Published on: September 29, 2017

20.2K

Area of Science:

  • Biomaterials Science
  • Stem Cell Biology
  • Tissue Engineering

Background:

  • Standard 2D cell cultures do not provide a physiologically relevant micro-environment for human mesenchymal stem cells (hMSCs).
  • hMSCs cultured in 2D lose their differentiation capacity and clinical potential.
  • A 3D micro-environment is crucial for maintaining hMSC phenotype and function.

Purpose of the Study:

  • To develop a 3D alginate hydrogel mimicking bone marrow's mechanical properties (<5 kPa) and functionalized with Arg-Gly-Asp (RGD).
  • To investigate the effect of this 3D micro-environment on hMSC cell cycle, proliferation, and quiescence markers.
  • To elucidate the molecular pathways, including mTORC1 signaling, involved in 3D-induced hMSC quiescence.

Main Methods:

  • Fabrication of 3D alginate hydrogels functionalized with RGD, with mechanical stiffness <5 kPa.
  • Culture of hMSCs in 3D hydrogels and comparison with 2D cultures.
  • Cell cycle analysis using flow cytometry and proliferation assessment via 5-Ethynyl-2'-deoxyuridine (EdU) staining.
  • Gene and protein expression analysis of quiescence markers (EZH1, FoxO3, p27) and mTORC1 signaling pathway.

Main Results:

  • hMSCs cultured in 3D RGD-functionalized hydrogels exhibited cell cycle arrest in G1 phase and became non-proliferative.
  • Quiescence in 3D cultures was associated with upregulated EZH1, FoxO3, and p27 expression compared to 2D cultures.
  • This quiescent phenotype was exclusive to 3D cultures and independent of RGD concentration or adhesion chemistry.
  • Downregulation of mTORC1 signaling was observed in 3D cultures, suggesting its role in inducing deep quiescence.

Conclusions:

  • 3D alginate hydrogels mimicking bone marrow stiffness and functionalized with RGD can induce a stable quiescent state in hMSCs.
  • The observed quiescence is linked to specific molecular pathways, including EZH1, FoxO3, p27 upregulation and mTORC1 downregulation.
  • These findings provide a better understanding of hMSC quiescence mechanisms in a 3D environment, crucial for advancing hMSC-based cell therapies.