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Related Concept Videos

Mesenchymal Stem Cells01:19

Mesenchymal Stem Cells

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Mesenchymal stem cells (MSCs) are adult stem cells that can differentiate into most connective tissue cell types, except for hematopoietic cells, depending upon the source of MSCs. For example, bone-marrow-derived MSCs (BM-MSCs) can differentiate into osteocytes, hepatocytes, and pancreatic and neuronal cells. MSCs can be isolated from various sources such as bone marrow, placenta, adipose tissue, teeth, and Wharton’s jelly, a gelatinous substance in the umbilical cord. The ease of their...
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Updated: Oct 9, 2025

Construction of a Multilayered Mesenchymal Stem Cell Sheet with a 3D Dynamic Culture System
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Construction of a Multilayered Mesenchymal Stem Cell Sheet with a 3D Dynamic Culture System

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Enhancing chondrogenic potential via mesenchymal stem cell sheet multilayering.

Hallie Thorp1,2, Kyungsook Kim1, Sophia Bou-Ghannam1,2

  • 1Cell Sheet Tissue Engineering Center (CSTEC), Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT, USA.

Regenerative Therapy
|December 20, 2021
PubMed
Summary

Multilayering mesenchymal stem cell (MSC) sheets enhances chondrogenesis up to a certain thickness. Exceeding this threshold reduces cartilage formation, indicating thickness is key for tissue engineering articular cartilage.

Keywords:
Cell sheet technologyCellular interactionsChondrogenic differentiationScaffold-freeTissue engineering

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Area of Science:

  • Tissue Engineering
  • Regenerative Medicine
  • Biomaterials Science

Background:

  • Mesenchymal stem cells (MSCs) are utilized in tissue engineering for cartilage repair due to their chondrogenic potential.
  • Cell sheet technology offers a scaffold-free, three-dimensional (3D) culture method for fabricating cartilage constructs in vitro.
  • Previous studies demonstrated the in vitro chondrogenic capacity of 3D MSC sheets.

Purpose of the Study:

  • To investigate the impact of cell sheet multilayering on the fabrication of MSC-derived hyaline-like cartilage constructs.
  • To assess structural and biochemical changes during chondrogenesis in multilayered MSC constructs.
  • To determine the optimal construct thickness for in vitro chondrogenesis using cell sheet multilayering.

Main Methods:

  • Preparation of multilayered MSC sheets.
  • Assessment of structural and biochemical properties throughout chondrogenesis.
  • Analysis of cellular interactions (N-cadherin, connexin 43, integrin β-1) and chondrogenic gene expression (Sox9, aggrecan, type II collagen).

Main Results:

  • MSC multilayering increased construct thickness and 3D cellular interactions.
  • Constructs of 25 μm (2-layer) showed enhanced chondrogenic interactions and differentiation compared to 14 μm (1-layer).
  • A thickness threshold (33 μm, 3-layer) was identified, leading to reduced chondrogenesis, gene expression, and proteoglycan accumulation.

Conclusions:

  • Cell sheet multilayering is a viable strategy for controlling construct thickness in MSC-based cartilage engineering.
  • Optimizing construct thickness is crucial for maximizing in vitro chondrogenesis and extracellular matrix deposition.
  • This approach holds promise for developing engineered cartilage for articular cartilage regeneration.