Fast isolation and expansion of multipotent cells from adipose tissue based on chitosan-selected primary culture

Huang, Guo Shiang; Tseng, Ting; Dai, Niann Tzyy; Fu, Keng Yen; Dai, Lien Guo; Hsu, Shan‐hui

doi:10.1016/j.biomaterials.2015.07.003

Cited by 8 publications

(3 citation statements)

References 51 publications

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“…It is a polysaccharide made of amino sugars and is technically widely used because of its biocompatibility. [6][7][8] Chitosan is composed of randomly distributed b-(1,4) linked D-glucosamine and N-acetyl-glucosamine (shown in Figure 1(a)). Interestingly, its chemical structure shares some characteristics with the chondroitin sulfate in AC (see Figure 1(b)), a type of sulfated glycosaminoglycans composed of repeating disaccharide structures of N-acetyl-galactosamine and glucuronic acids.…”

Section: Introductionmentioning

confidence: 99%

Genipin crosslinked chitosan/PEO nanofibrous scaffolds exhibiting an improved microenvironment for the regeneration of articular cartilage

et al. 2021

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Towards optimizing the growth of extracellular matrix to produce repair cartilage for healing articular cartilage (AC) defects in joints, scaffold-based tissue engineering approaches have recently become a focus of clinical research. Scaffold-based approaches by electrospinning aim to support the differentiation of chondrocytes by providing an ultrastructure similar to the fibrillar meshwork in native cartilage. In a first step, we demonstrate how the blending of chitosan with poly(ethylene oxide) (PEO) allows concentrated chitosan solution to become electrospinnable. The chitosan-based scaffolds share the chemical structure and characteristics of glycosaminoglycans, which are important structural components of the cartilage extracellular matrix. Electrospinning produced nanofibrils of ∼100 nm thickness that are closely mimicking the size of collagen fibrils in human AC. The polymer scaffolds were stabilized in physiological conditions and their stiffness was tuned by introducing the biocompatible natural crosslinker genipin. We produced scaffolds that were crosslinked with 1.0% genipin to obtain values of stiffness that were in between the stiffness of the superficial zone human AC of 600 ± 150 kPa and deep zone AC of 1854 ± 483 kPa, whereas the stiffness of 1.5% genipin crosslinked scaffold was similar to the stiffness of deep zone AC. The scaffolds were degradable, which was indicated by changes in the fibril structure and a decrease in the scaffold stiffness after seven months. Histological and immunohistochemical analysis after three weeks of culture with human articular chondrocytes (HACs) showed a cell viability of over 90% on the scaffolds and new extracellular matrix deposited on the scaffolds.

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Section: Introductionmentioning

confidence: 99%

Genipin crosslinked chitosan/PEO nanofibrous scaffolds exhibiting an improved microenvironment for the regeneration of articular cartilage

et al. 2021

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“…Adipose-derived MSCs isolated by CS substrates displayed higher proliferation rates and stemness, as compared to those collected by conventional plastic adherence approach. The expression of several stemness markers, such as Oct4, Sox2, and Nanog, as well as MSC marker, CD271 [54], were well maintained in the MSC spheroids generated by CS substrates [55].…”

Section: Spheroid Culture Of Mscsmentioning

confidence: 94%

Cellular Spheroids of Mesenchymal Stem Cells and Their Perspectives in Future Healthcare

2019

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Intrinsic cellular properties of several types of cells are dramatically altered as the culture condition shifts from two-dimensional (2D) to three-dimensional (3D) environment. Currently, several lines of evidence have demonstrated the therapeutic potential of mesenchymal stem cells (MSCs) in regenerative medicine. MSCs not only replenish the lost cells, they also promote the regeneration of impaired tissues by modulating the immune responses. Following the development of 3D cell culture, the enhanced therapeutic efficacy of spheroid-forming MSCs have been identified in several animal disease models by promoting differentiation or trophic factor secretion, as compared to planar-cultured MSCs. Due to the complicated and multifunctional applications in the medical field, MSCs are recently named as medicinal signaling cells. In this review, we summarize the predominant differences of cell-environment interactions for the MSC spheroids formed by chitosan-based substrates and other scaffold-free approaches. Furthermore, several important physical and chemical factors affecting cell behaviors in the cell spheroids are discussed. Currently, the understanding of MSCs spheroid interactions is continuously expanding. Overall, this article aims to review the broad advantages and perspectives of MSC spheroids in regenerative medicine and in future healthcare. Three-Dimensional (3D) Cell Culture SystemsCell-cell and cell-environment interactions cooperatively determine the physiological phenomena in vivo. Many cell growth mechanisms and signal transduction pathways occurring in cells have been unveiled by the traditional two-dimensional (2D) culture system. Plastic-based culture materials remain the predominant cell culture platforms to date, yet some concerns have been raised with the accumulating experimental evidence.A major drawback of cell culture on 2D plastic plates is the planar cell-cell interaction that has the insufficient and inappropriate cell-environment crosstalk. In vivo, cells are grown and stacked within a three-dimensional (3D) space, and extracellular matrix (ECM) secreted by cells constitutes the supporting framework to further organize tissues and organs. Therefore, some important cell-cell and cell-environment interactions may be ignored in a planar culture platform. Primary cells, cancer cells, and stem cells have been cultured in a 3D environment, and many interesting findings have been uncovered. In general, the definition of 3D cell culture systems is that cells are cultured on/in the 3D scaffolds composed by natural or synthetic materials, or they are organized into cellular spheroids. With the continuing development of 3D cell culture, it is now generally agreed that the cell physiology and cell behavior are dramatically distinct for cells cultured within a 2D or 3D environment [1][2][3].Cancer cells are the most common cell types that have been investigated by 3D culture systems, and obvious change of gene expression as well as intracellular signaling in these 3D-cultured cancer cells have been ...

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“…cultures. As a result, 2D cell culture methods have the limitation that they do not accurately reflect the in vivo environment, and as the cells grow and proliferate, the unnatural 2D environment can also affect gene expression [6][7][8][9][10]. To overcome the limitations of these 2D cell culture methods, a three-dimensional (3D) cell culture method has recently been developed and used to grow cells in 3D and create an environment similar to the in vivo conditions [11,12].…”

Section: S1 Fig Shows the Differences Between 2d And 3d Cellmentioning

confidence: 99%

Testingin vitrotoxicity of nanoparticles in 3D cell culture with various extracellular matrix scaffold

Bae

et al. 2021

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Nanomaterials are used in a variety of fields and toxicity assessment is paramount for their development and application. Although most toxicity assessments have been performed in 2D (2-Dimensional) cell culture, the inability to adequately replicate the in vivo environment and toxicity is a limitation. To overcome the limitation, a 3D (3-Dimensional) cell culture method has been developed to make an environment closer to an in vivo system. In this study, 20 nm SiO2 nanoparticles were dispersed in serum-containing (SC) and serum-free (SF) media to compare 2D cell culture and 3D cell culture toxicity. The cells were subjected to a 3D cell culture method in which HepG2, a human-derived liver cancer cell line, was mixed on a scaffold. We found that nanoparticles induced toxicity in 2D cell culture, but toxicity was not observed in 3D cell culture similar to in vivo environment. However, differences in toxicity were observed between the three types of scaffolds in the absence of serum as the number of cells decreased.

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Fast isolation and expansion of multipotent cells from adipose tissue based on chitosan-selected primary culture

Cited by 8 publications

References 51 publications

Genipin crosslinked chitosan/PEO nanofibrous scaffolds exhibiting an improved microenvironment for the regeneration of articular cartilage

Genipin crosslinked chitosan/PEO nanofibrous scaffolds exhibiting an improved microenvironment for the regeneration of articular cartilage

Cellular Spheroids of Mesenchymal Stem Cells and Their Perspectives in Future Healthcare

Testingin vitrotoxicity of nanoparticles in 3D cell culture with various extracellular matrix scaffold

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