Modulating and modeling aggregation of cell-seeded microcarriers in stirred culture system for macrotissue engineering

Yang, Mei; Luo, Hao; Tang, Qiang; Ye, Zhaoyang; Zhou, Yan; Tan, Wen‐Song

doi:10.1016/j.jbiotec.2010.09.953

Cited by 27 publications

(48 citation statements)

References 36 publications

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“…These micro-aggregates with predefined shape and dimension could be transplanted at the sites of meniscus defects [Katagiri et al, 2013] or used as building blocks for meniscal repair. Assembly of these building blocks into larger tissues might be obtained by self-assembly [Wolf et al, 2008] or by the use of appropriate biomaterials, creating the possibility to assemble the micro-aggregates in a more direct way [Mironov et al, 2009;Mei et al, 2010] and thereby demonstrating their potential in cell-based tissue engineering strategies. The objective of this study was (1) to investigate the changes that occur during monolayer expansion of porcine fibrochondrocytes, (2) to investigate if uniform fibrochondrocyte micro-aggregates with desirable phenotype and tissue homogeneity could be produced with our microwell platform, and (3) since fibrochondrocytes are exposed to a hypoxic environment [Malda et al, 2003b] in vivo the effect of hypoxia on the matrix quality of these generated micro-aggregates was investigated.…”

Section: Introductionmentioning

confidence: 99%

Redifferentiation of High-Throughput Generated Fibrochondrocyte Micro-Aggregates: Impact of Low Oxygen Tension

Berneel¹,

Philips²,

Declercq³

et al. 2016

Cells Tissues Organs

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In meniscus tissue engineering strategies, enhancing the matrix quality of the neomeniscal tissue is important. When the differentiated phenotype of fibrochondrocytes is lost, the quality of the matrix becomes compromised. The objective of this study was to produce uniform fibrochondrocyte micro-aggregates with desirable phenotype and tissue homogeneity in large quantities using a simple and reproducible method. Furthermore, we investigated if hypoxia could enhance the matrix quality. Porcine fibrochondrocytes were expanded at 21% oxygen until passage 3 (P3) and a gene expression profile was determined. P3 fibrochondrocytes were cultivated in chondrogenic medium at 5 and 21% oxygen in high-throughput agarose chips containing 2,865 microwells 200 µm in diameter. Evaluation included live/dead staining, histological examination, immunohistochemistry, dimethylmethylene blue assay and real-time reverse transcriptase quantitative polymerase chain reaction of the micro-aggregates. Gene expression analysis showed a drastic decline in collagen II and high expression of collagen I during monolayer culture. After 4 days, uniform and stable micro-aggregates could be produced. The redifferentiation and matrix quality of the hypoxic cultured micro-aggregates were enhanced relative to the normoxic cultures. Sulfated glycosaminoglycan synthesis was significantly higher, and collagen II expression and the collagen II/collagen I ratio were significantly upregulated in the hypoxic cultures. High-throughput production of uniform microtissues holds promise for the generation of larger-scale tissue engineering constructs or optimization of redifferentiation mechanisms for clinical applications.

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Section: Introductionmentioning

confidence: 99%

Redifferentiation of High-Throughput Generated Fibrochondrocyte Micro-Aggregates: Impact of Low Oxygen Tension

Berneel¹,

Philips²,

Declercq³

et al. 2016

Cells Tissues Organs

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show abstract

“…To circumvent mass transfer limitation encountered in conventional top-down methodology, we previously proposed so-called "microtisse assembly technique" for fabricating macroscopic tissue constructs [8,14]. This procedure essentially includes two steps as illustrated in Figure 1: cells are seeded onto microcarriers and cultivated in spinner flasks to prepare Accepted Article Biotechnology Journal microtissues, which are then subjected to perfusion culture for assembling into macrotissues.…”

Section: Discussionmentioning

confidence: 99%

“…This procedure essentially includes two steps as illustrated in Figure 1: cells are seeded onto microcarriers and cultivated in spinner flasks to prepare Accepted Article Biotechnology Journal microtissues, which are then subjected to perfusion culture for assembling into macrotissues. Our previous reports had concerned the fabrication of microtissues and also demonstrated successful preparation of a macroscopic bone construct from hMSCs [8,14]. Among different strategies employed to assemble microtissues in literature [7,23,24], perfusion culture is able to promote interstitial flow and exert shearing force on cells and may favor cell growth and differentiation and tissue maturation [13,16,19,25].…”

Section: Discussionmentioning

confidence: 99%

“…It should be noted that HepG2 cells tended to detach from microcarriers during culture, leading to lower cell density on Cytopore-2 than HFs and hMSCs (data not shown). In addition, on CultiSpher S, cells tended to remain on the outer surface, which potentially favored the formation of cell bridging between microcarriers, thus promoting the aggregation for cell-laden CultiSpher S [8,29]. HepG2 cells-laden microcarriers always formed smaller aggregates than other two cell types, suggesting that cell type also affected the aggregation Accepted Article Biotechnology Journal process.…”

Section: Discussionmentioning

confidence: 99%

“…Previously, we had proposed "microtissue assembling technique" by assembling cell-laden microcarriers to fabricate macrotissues with large dimensions through perfusion culture, which was expected to surpass the limit of mass transfer encounted in conventional top-down tissue engineering [8].…”

Section: Introductionmentioning

confidence: 99%

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Perfusion culture‐induced template‐assisted assembling of cell‐laden microcarriers is a promising route for fabricating macrotissues

Xiu¹,

Jiao²,

Zhang³

et al. 2014

Biotechnology Journal

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Mass transfer limitation in conventional top-down tissue engineering makes it impossible to fabricate large size viable tissue replacements. In the present study, we aimed at performing a systemic investigation of the assembling process in perfusion culture for fabricating centimeter-scale macrotissues from cell-laden microcarriers following a bottom-up modular approach. Cells (human fibroblasts, human mesenchymal stem cells, or HepG2 cells) were seeded onto microcarriers (Cytopore-2 or CultiSpher S) in spinner flasks and cultured for 14 days and subsequently transferred to a perfusion chamber for assembling. It was found that growth of different cells on different microcarriers varied and aggregation of cell-laden microcarriers was favored with CultiSpher S. After perfusion culture for 14 days, while all microtissues could assemble into integral macrotissues, macrotissues of HepG2 cells were structurally most inferior and the assembling of cell-laden CultiSpher S led to a significant shrinkage. By designing perfusion chamber and using agar-based templates, tubular, disc, and alphabetic letter-shaped macrotissues could be easily fabricated, suggesting template-assisted assembling. Importantly, it was revealed that there existed both optimal perfusion culture time (21 days) and packing condition (1/4 compression) for the assembling of microtissues. This study lays a solid foundation for future applications of this microtissue assembling technique in tissue engineering.

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Fabrication of viable and functional pre‐vascularized modular bone tissues by coculturing MSCs and HUVECs on microcarriers in spinner flasks

Zhang

Zhou

et al. 2017

Biotechnology Journal

Self Cite

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Modulating and modeling aggregation of cell-seeded microcarriers in stirred culture system for macrotissue engineering

Cited by 27 publications

References 36 publications

Redifferentiation of High-Throughput Generated Fibrochondrocyte Micro-Aggregates: Impact of Low Oxygen Tension

Redifferentiation of High-Throughput Generated Fibrochondrocyte Micro-Aggregates: Impact of Low Oxygen Tension

Perfusion culture‐induced template‐assisted assembling of cell‐laden microcarriers is a promising route for fabricating macrotissues

Fabrication of viable and functional pre‐vascularized modular bone tissues by coculturing MSCs and HUVECs on microcarriers in spinner flasks

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