A novel 3-D model for cell culture and tissue engineering

Zhang, Xulang; Xie, Yizhen; Koh, Chee Guan; Lee, L. James

doi:10.1007/s10544-009-9294-8

Cited by 22 publications

(18 citation statements)

References 27 publications

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“…Beyond differences in mass transport kinetics, studies have also indicated that culturing cell in 3-D matrix instead of 2-D surfaces lead to alteration in cell phenotypes and function, including differentiation 21, 22 , proliferation 23 and survival 24 . For example, many tumor cells display reduced chemosensitivity in 3-D cultures 25 .…”

Section: Resultsmentioning

confidence: 99%

Microscale 3D Collagen Cell Culture Assays in Conventional Flat-Bottom 384-Well Plates

et al. 2015

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Three-dimensional culture systems such as cell-laden hydrogels are superior to standard 2-D monolayer cultures for many drug-screening applications. However, their adoption in high throughput screening (HTS) have been lagging, in part due to the difficulty of incorporating these culture formats into existing robotic liquid handling and imaging infrastructures. Dispensing cell-laden pre-polymer solutions into 2-D well-plates is a potential solution, but typically requires large volumes of reagents to avoid evaporation during polymerization, which increases cost, makes drug penetration variable and imaging complex. Here we describe a technique to efficiently produce 3-D ‘microgels’ using automated liquid handling systems and standard, non-patterned, flat-bottomed, 384-well plates. Sub-millimeter-diameter, cell-laden collagen gels are deposited on the bottom of ~2.5 mm-diameter microwell with no concerns over evaporation and meniscus effects at the edges of wells, using aqueous two-phase system patterning. The microscale cell-laden collagen-gel constructs are readily imaged and readily penetrated by drugs. Cytotoxicity of chemotherapeutics were monitored by bioluminescence and demonstrates that 3-D cultures confer chemoresistance, as compared to similar 2-D culture. This data hence, demonstrates the importance of culturing cells in 3-D to obtain realistic cellular responses. Overall, this system provided a simple and inexpensive method for integrating 3-D culture capability into existing HTS infrastructure.

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Section: Resultsmentioning

confidence: 99%

Microscale 3D Collagen Cell Culture Assays in Conventional Flat-Bottom 384-Well Plates

et al. 2015

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“…After 16 days of cultivation in static dish, the physiological density of live cells was found only within the top 128 mm of construct thickness. A tissue fabricated from suspended cells in microcapsules for cell-based drug delivery and tumor models shows its critical size (approximately 200 -300 mm) [22]. In these methods, oxygen concentration and cell viability decrease with the distance from the construct surface.…”

Section: Discussionmentioning

confidence: 99%

Thickness limitation and cell viability of multi-layered cell sheets and overcoming the diffusion limit by a porous-membrane culture insert

Sekine¹,

Haraguchi²

2011

J Biochip Tissue chip

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A major challenge of tissue engineering is the creation of three-dimensional (3D) and functional tissues. However, the tissue ischemic environments make the production of thicker 3D tissues difficult. For evaluating the thickness limitation of tissue, the cell viability and metabolism of a novel in vitro 3D tissue model, which is composed of multilayered cell sheets, were investigated. Human endometrial-derived mesenchymal cells (EMC) were cultured on temperature-responsive culture dishes. Confluently cultured EMCs were harvested as an intact contiguous cell sheet only by lowering temperature. The obtained cell sheets were successfully layered into 3D cell-dense tissues and cultured in vitro for 7 days. Glucose consumption and lactate production in the culture media increased in accordance with the number of layers (single to triple). Histological analyses and cell viability assays showed that viable tissues were found in single-to triple-layered cell sheets and damaged tissues in over quadruple layers. The results concluded that the thickness limitation of layered cell sheets was approximately 40 mm, which was the thickness of triple-layered cell sheets. Importantly, when multi-layered cell sheets were cultured on porous membranes, the adhesion among cell sheets and cell viability were improved, resulting in successful fabricating thicker tissues (~100 mm) than that on normal culture dishes. The metabolic analyses showed that multi-layered cell sheets rely on their anaerobic metabolism, indicating that supplying nutrients rather than oxygen through both upper and bottom tissue surfaces improved the cell viability in 3D tissues. ` `Citation: Sekine W, Haraguchi Y, Shimizu T, Umezawa A, Okano T (2011) Thickness limitation and cell viability of multi-layered cell sheets and overcoming the diffusion limit by a porous-membrane culture insert. J Biochip Tissue chip S1:007.

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“…Factors used for stem cell differentiation include signals from blood vessels such as vascular endothelial growth factor A (VEGF-A) and fetal soluble factors, which play an important role in the pancreatic differentiation of embryonic stem cells. [46][47][48][49][50] Additionally, undifferentiated embryonic stem cells are genetically engineered with b cell genes such as Nkx6.1 to obtain insulin-secreting cells. …”

Section: Embryonic Stem Cellsmentioning

confidence: 99%

Islet Encapsulation: Strategies to Enhance Islet Cell Functions

Beck¹,

Angus²,

Madsen³

et al. 2007

Tissue Engineering

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Diabetes is one of the most prevalent, costly, and debilitating diseases in the world. Although traditional insulin therapy has alleviated the short-term effects, long-term complications are ubiquitous and harmful. For these reasons, alternative treatment options are being developed. This review investigates one appealing area: cell replacement using encapsulated islets. Encapsulation materials, encapsulation methods, and cell sources are presented and discussed. In addition, the major factors that currently limit cell viability and functionality are reviewed, and strategies to overcome these limitations are examined. This review is designed to introduce the reader to cell replacement therapy and cell and tissue encapsulation, especially as it applies to diabetes.

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A novel 3-D model for cell culture and tissue engineering

Cited by 22 publications

References 27 publications

Microscale 3D Collagen Cell Culture Assays in Conventional Flat-Bottom 384-Well Plates

Microscale 3D Collagen Cell Culture Assays in Conventional Flat-Bottom 384-Well Plates

Thickness limitation and cell viability of multi-layered cell sheets and overcoming the diffusion limit by a porous-membrane culture insert

Islet Encapsulation: Strategies to Enhance Islet Cell Functions

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