Tissue engineering: a new frontier in physiological genomics

Petersen, Matthew C.; Lazar, Jozef; Jacob, Howard J.; Wakatsuki, Toshiyuki

doi:10.1152/physiolgenomics.00165.2007

Cited by 11 publications

(6 citation statements)

References 36 publications

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“…A successful comparison of gene expressions of five calcium‐handling proteins between a neonatal heart and various culturing conditions was achieved. It was concluded that culturing the cells in a three‐dimensional environment produces tissues that more closely resemble native cardiac tissue (29–31). There still needs to be a lot more research done in order to develop the perfect culturing conditions which will provide the correct and adequate amount of nutrients to each cell.…”

Section: Discussionmentioning

confidence: 99%

Changes in Gene Expression During the Formation of Bioengineered Heart Muscle

Khait

Birla

2009

Artificial Organs

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A three-dimensional bioengineered heart muscle (BEHM) construct model had been previously developed, exhibiting contractile forces up to 800 microN. The interest of this study was to determine gene expression levels of biologic markers involved in calcium-handling between BEHM, cell monolayer, and neonatal heart. Cardiac cells were isolated from one litter of F344 rats and organized into groups (n = 5): 4-, 7-, 10-day BEHM and cell monolayer; BEHM was evaluated for cell viability and contractility. Groups were then analyzed for mRNA expression of calcium-handling proteins: myosin heavy chain (MHC) alpha and beta, Sarcoplasmic reticulum Ca++ ATPase (SERCA) 2, phospholamban (PBL), and ryanodine receptor. BEHM exhibited electrically stimulated active force (208 +/- 12 microN day 4, 361 +/- 22 microN day 7, and 344 +/- 29 microN day 10) and no decrease in cell number. Real-time polymerase chain reaction (PCR) showed an increase in gene expression of all calcium-handling proteins in BEHM at 7 and 10 days compared with monolayers, for example, comparing BEHM to monolayer (7 and 10 days, respectively), MHC-alpha: 2600-fold increase and a 100-fold increase; MHC-beta: 70-fold increase at 10 days; ryanodine receptor: 74-fold increase at 10 days; SERCA: 19-fold increase and sixfold increase; PBL: 158-fold increase and 24-fold increase. It was concluded that a three-dimensional environment is a better culturing condition of cardiac cells than a monolayer. Also, BEHM constructs demonstrated a high similarity to a native myocardium, and is, thus, a good starting foundation for engineered heart muscle.

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Section: Discussionmentioning

confidence: 99%

Changes in Gene Expression During the Formation of Bioengineered Heart Muscle

Khait

Birla

2009

Artificial Organs

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“…Cell culture techniques must be adapted over scaffolds with 3D architecture, designed in scales relevant to tissue natural physiologic processes [10]. The main goal of using tissue engineering in biomedical research is to combine traditional two-dimensional (2D) cell culture and in vivo conditions, submitting cultured cells to an environment that better represent the complex 3D structure of the native tissue, where cells grow surrounded by the extracellular matrix and neighboring cells in all three dimensions [11]. Therefore, we aim to perform comparative biocompatibility in vitro studies, evaluating cell cultures both in monolayer and in an organized 3D structure in order to characterize the limitations and advantages of each method, considering two different types of cells cultures.…”

Section: Introductionmentioning

confidence: 99%

Three-Dimensional Systems in Polybutylcyanoacrylate Nanoparticles Safety Evaluation

Santos

Tiago

Maria‐Engler

et al. 2015

International Journal of Polymeric Materials and Polymeric Biom

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It is generally accepted that three-dimensional (3D) cell culture systems better represent cell physiology and morphology than two-dimensional (monolayer) cell cultures. The aim of this work was to develop a 3D model that could efficiently evaluate polymeric materials biocompatibility using polybutylcyanoacrylate nanoparticles (PBCA-NP). Normal human fibroblasts and melanoma cells were cultured both in monolayer and on type I collagen matrix. Trypan blue and MTT assays were used to evaluate PBCA-NP cytotoxicity, which was shown to be dose-and time-dependent to all cell types. These results were confirmed by cell death analysis with flow cytometry. There is evidence that treatment with PBCA-NP in concentrations close to the IC 50 inhibit autophagy in fibroblasts and melanoma cells when cultured in monolayer, but this response could not be observed in the 3D system. These findings strongly suggest that PBCA-NP can modify cell response depending on the concentration used and the conditions of cell culture.

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“…Therefore, they have been commonly used to examine cancer cell biology and develop new drugs. However, tumor cells grown in 2D culture systems differ from those grown in vivo; they have different morphology, cell-to-cell or cell-to-matrix adhesions, cellular differentiation patterns, and cellular behaviors (3,4).…”

Section: Introductionmentioning

confidence: 99%

Morphological and cytoskeletal alterations of nervous system tumor cells with different culturing methods

Matsuda

Kawamoto²,

Teduka³

et al. 2011

Int J Oncol

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Abstract. Cell culture is one of the most important methods of research in molecular and cellular biology, and various culture systems have been developed, including two-dimensional (2D), three-dimensional (3D) and floating culture systems. In the present study, we examined morphological changes and different expression patterns of cytoskeletal proteins in three different types of nervous system tumor cells grown in 2D, 3D and floating cell cultures. A172, KG-1-C and IMR-32 cells showed marked morphological changes, depending on the cell culture methods. F-actin expression was clearly observed at the level of the cells nearest the plate surface in 2D and 3D cultures. On the other hand, expression of F-actin was weak in the floating culture system. α-tubulin was detected in the cytoplasm of cells in 2D culture, but in floating and 3D cultures, α-tubulin was expressed in the peripheral regions of spheres and spheroids. In conclusion, this study demonstrated that nervous system tumor cells showed different alterations in morphology, and different cytoskeletal protein expression patterns, depending on the culture methods. IntroductionWith the aim of mimicking in vivo situations or particular pathological conditions, various culture systems have been developed, including two-dimensional (2D), three-dimensional (3D), and floating culture systems (1,2). Traditional tumor cell culture studies with 2D cultures have the advantages of ease and economy. Therefore, they have been commonly used to examine cancer cell biology and develop new drugs. However, tumor cells grown in 2D culture systems differ from those grown in vivo; they have different morphology, cell-to-cell or cell-to-matrix adhesions, cellular differentiation patterns, and cellular behaviors (3,4).On the other hand, 3D cell culture systems were expected to mimic in vivo environments (5-7). Many types of 3D culture systems have been reported, including spheroid cultures, ex vivo cultures (8), multilayered postconfluent cell cultures (9), cells grown in an extracellular matrix gel (10), and co-cultures with fibroblasts or vascular endothelium (11). The recently developed NanoCulture plate, which has a microsquare pattern on the bottom of the plate, allows the formation of 3D spheroids without the need of any matrix or scaffold. Recently, we reported that several kinds of pancreatic cancer cell lines formed cell spheroids in NanoCulture plates (12). These cells showed alterations in the expression levels and localization of cytoskeletal proteins. The 3D spheroidal culture system with the NanoCulture plate is a useful method for cell imaging with phase contrast microscopy and confocal microscopy.Furthermore, floating cultures in plates with a low-attachment surface permit the formation of spheres of several kinds of cells. It has been reported that neural stem cells and tumor stem cells grow in floating spheres in serum-free culture medium supplemented with epidermal growth factor (EGF) and/or basic fibroblast growth factor (bFGF) (13,14). Therefore, the floating ...

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Tissue engineering: a new frontier in physiological genomics

Cited by 11 publications

References 36 publications

Changes in Gene Expression During the Formation of Bioengineered Heart Muscle

Changes in Gene Expression During the Formation of Bioengineered Heart Muscle

Three-Dimensional Systems in Polybutylcyanoacrylate Nanoparticles Safety Evaluation

Morphological and cytoskeletal alterations of nervous system tumor cells with different culturing methods

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