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Human Pluripotent Stem Cell Culture on Polyvinyl Alcohol-Co-Itaconic Acid Hydrogels with Varying Stiffness Under Xeno-Free Conditions
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Dynamic and self-biodegradable polysaccharide hydrogel stores embryonic stem cell construct under ambient condition.

Kuan Yang1, Wei Wei1, Li Ting Gao1

  • 1College of Bioresources Chemical and Materials Engineering, National Demonstration Center for Experimental Light Chemistry Engineering Education, Shaanxi University of Science and Technology, Xi'an, China.

Frontiers in Bioengineering and Biotechnology
|May 30, 2023
PubMed
Summary

We developed a novel dynamic hydrogel construct for storing and transporting mouse embryonic stem cells (mESCs). This method ensures high survival rates and preserves pluripotency for "off-the-shelf" stem cell applications.

Keywords:
ambient conditiondynamic polysaccharide hydrogelembryonic stem cell constructself-biodegradationstorage

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Area of Science:

  • Biomaterials Science
  • Stem Cell Biology
  • Tissue Engineering

Background:

  • Embryonic stem cells (ESCs) require specific microenvironments for storage and transport.
  • Current methods face challenges in maintaining cell viability and function during transit.
  • The need for readily available, viable stem cells for research and therapeutic applications is increasing.

Purpose of the Study:

  • To develop a novel method for storing and transporting ESCs under ambient conditions.
  • To create an "off-the-shelf" solution for stem cell delivery using a dynamic hydrogel.
  • To assess the viability, pluripotency, and long-term culture capacity of ESCs stored and transported in a hydrogel construct.

Main Methods:

  • Mouse ESCs (mESCs) were encapsulated within a self-biodegradable, dynamic, polysaccharide-based hydrogel to form ESCs-dynamic hydrogel constructs (CDHC).
  • CDHC were stored under sterile, hermetic conditions for 6 days, including transport simulation.
  • Post-transport, mESCs were released from the hydrogel and cultured for 15 generations to assess recovery and pluripotency.

Main Results:

  • Large, compact mESC colonies maintained a 90% survival rate and pluripotency after storage and simulated transport.
  • Encapsulated stem cells were automatically released from the self-biodegradable hydrogel upon arrival.
  • Retrieved mESCs demonstrated restored colony-forming capacity and pluripotency markers at both protein and mRNA levels after 15 generations of subculture.

Conclusions:

  • The dynamic, self-biodegradable hydrogel offers a simple, cost-effective method for ambient storage and transport of ESCs.
  • This approach facilitates "off-the-shelf" availability of viable stem cells.
  • The CDHC technology supports widespread applications of stem cells in research and potentially clinical settings.