Effect of Degree of Deacetylation of Chitosan/Chitin on Human Neural Stem Cell Culture

Chang, Fei‐Chien; Zhou, Yang; James, Matthew Michael; Zareie, Hadi M; Ando, Yumiko; Yang, Jihui; Zhang, Miqin

doi:10.1002/mabi.202200389

Cited by 5 publications

(2 citation statements)

References 82 publications

(132 reference statements)

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“…The localized compression, tension, and nano- or microscale features of the substrate play a crucial role in regulating cell fate through cell transmembrane receptors, ultimately influencing cytoskeleton and cell morphology. In our previous study, we investigated the impact of different degrees of deacetylation on the mechanical properties of chitosan films [ 67 ]. We found that a higher degree of deacetylation led to a decrease in the elastic modulus of the films.…”

Section: Resultsmentioning

confidence: 99%

Chitosan Scaffolds as Microcarriers for Dynamic Culture of Human Neural Stem Cells

et al. 2023

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Human neural stem cells (hNSCs) possess remarkable potential for regenerative medicine in the treatment of presently incurable diseases. However, a key challenge lies in producing sufficient quantities of hNSCs, which is necessary for effective treatment. Dynamic culture systems are recognized as a powerful approach to producing large quantities of hNSCs required, where microcarriers play a critical role in supporting cell expansion. Nevertheless, the currently available microcarriers have limitations, including a lack of appropriate surface chemistry to promote cell adhesion, inadequate mechanical properties to protect cells from dynamic forces, and poor suitability for mass production. Here, we present the development of three-dimensional (3D) chitosan scaffolds as microcarriers for hNSC expansion under defined conditions in bioreactors. We demonstrate that chitosan scaffolds with a concentration of 4 wt% (4CS scaffolds) exhibit desirable microstructural characteristics and mechanical properties suited for hNSC expansion. Furthermore, they could also withstand degradation in dynamic conditions. The 4CS scaffold condition yields optimal metabolic activity, cell adhesion, and protein expression, enabling sustained hNSC expansion for up to three weeks in a dynamic culture. Our study introduces an effective microcarrier approach for prolonged expansion of hNSCs, which has the potential for mass production in a three-dimensional setting.

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Section: Resultsmentioning

confidence: 99%

Chitosan Scaffolds as Microcarriers for Dynamic Culture of Human Neural Stem Cells

et al. 2023

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“…In our earlier investigations, we have demonstrated that chitosan films with a higher degree of deacetylation (DD) exhibit the capacity to enhance human neural stem cell proliferation and sustain pluripotency. 37 In this study, 3D chitosan scaffolds were made out of medical grade chitosan (DD = 98.38%) using a freeze-drying technique to achieve a highly porous architecture. The architecture of these scaffolds was first examined in both their dry and hydrated states through scanning electron microscopy (SEM) and optical microscopy.…”

Section: Chitosan Scaffold Fabrication and Characterizationmentioning

confidence: 99%

A Chitosan Scaffold Supports the Enhanced and Prolonged Differentiation of HiPSCs into Nucleus Pulposus-like Cells

Tang,

Zhou,

Zhang

2024

ACS Appl. Mater. Interfaces

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Intervertebral disc degeneration (IDD) is a progressive condition and stands as one of the primary causes of low back pain. Cell therapy that uses nucleus pulposus (NP)-like cells derived from human induced pluripotent stem cells (hiPSCs) holds great promise as a treatment for IDD. However, the conventional two-dimensional (2D) monolayer cultures oversimplify cell−cell interactions, leading to suboptimal differentiation efficiency and potential loss of phenotype. While three-dimensional (3D) culture systems like Matrigel improve hiPSC differentiation efficiency, they are limited by animal-derived materials for translation, poorly defined composition, short-term degradation, and high cost. In this study, we introduce a new 3D scaffold fabricated using medicalgrade chitosan with a high degree of deacetylation. The scaffold features a highly interconnected porous structure, near-neutral surface charge, and exceptional degradation stability, benefiting iPSC adhesion and proliferation. This scaffold remarkably enhances the differentiation efficiency and allows uninterrupted differentiation for up to 25 days without subculturing. Notably, cells differentiated on the chitosan scaffold exhibited increased cell survival rates and upregulated gene expression associated with extracellular matrix secretion under a chemically defined condition mimicking the challenging microenvironment of intervertebral discs. These characteristics qualify the chitosan scaffold-cell construct for direct implantation, serving as both a structural support and a cellular source for enhanced stem cell therapy for IDD.

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Human Neural Stem Cell Expansion in Natural Polymer Scaffolds Under Chemically Defined Condition

Chang,

James,

Zhou

et al. 2024

Advanced Biology

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The maintenance and expansion of human neural stem cells (hNSCs) in 3D tissue scaffolds is a promising strategy in producing cost‐effective hNSCs with quality and quantity applicable for clinical applications. A few biopolymers have been extensively used to fabricate 3D scaffolds, including hyaluronic acid, collagen, alginate, and chitosan, due to their bioactive nature and availability. However, these polymers are usually applied in combination with other biomolecules, leading to their responses difficult to ascribe to. Here, scaffolds made of chitosan, alginate, hyaluronic acid, or collagen, are explored for hNSC expansion under xeno‐free and chemically defined conditions and compared for hNSC multipotency maintenance. This study shows that the scaffolds made of pure chitosan support the highest adhesion and growth of hNSCs, yielding the most viable cells with NSC marker protein expression. In contrast, the presence of alginate, hyaluronic acid, or collagen induces differentiation toward immature neurons and astrocytes even in the maintenance medium and absence of differentiation factors. The cells in pure chitosan scaffolds preserve the level of transmembrane protein profile similar to that of standard culture. These findings point to the potential of using pure chitosan scaffolds as a base scaffolding material for hNSC expansion in 3D.

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Effect of Degree of Deacetylation of Chitosan/Chitin on Human Neural Stem Cell Culture

Cited by 5 publications

References 82 publications

Chitosan Scaffolds as Microcarriers for Dynamic Culture of Human Neural Stem Cells

Chitosan Scaffolds as Microcarriers for Dynamic Culture of Human Neural Stem Cells

A Chitosan Scaffold Supports the Enhanced and Prolonged Differentiation of HiPSCs into Nucleus Pulposus-like Cells

Human Neural Stem Cell Expansion in Natural Polymer Scaffolds Under Chemically Defined Condition

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