Engineering microenvironment for expansion of sensitive anchorage-dependent mammalian cells

Chia, Ser-Mien; Lin, Pao-Chun; Yin, Chao; Mao, Hai‐Quan; Leong, Kam W.; Xu, Xi; Goh, Cho-Hong; Ng, Mah-Lee; Yu, Hanry

doi:10.1016/j.jbiotec.2005.05.012

Cited by 8 publications

(8 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For collagen nano-fibre characterization, microcapsules were imaged immediately after they were formed using an Olympus Fluoview 500 confocal microscope in back-scattering mode using a 60 Â WLSM lens of NA 1.00 [21]. Optical sectioning of 2 mm was used to obtain a 3-D stack of the microcapsule.…”

Section: Imaging and Characterization Of Nano-fibres In Microcapsulementioning

confidence: 99%

“…Dual labelling of collagen with fluorescein isothiocynate (FITC) and terpolymer with 9-anthroylnitrile (9-AN) (Molecular Probes) were described previously [21,25]. The microcapsules were also processed for scanning electron microscopy and imaged with a JOEL 5600 LV as described previously [14].…”

Section: Imaging and Characterization Of Nano-fibres In Microcapsulementioning

confidence: 99%

“…Collagen methylation, which involves the esterification of the carboxyl groups on collagen, results in its net positive charge and reduced tendency to gel because of charge repulsion [21,26]. The complex coacervation between the oppositely charged methylated collagen and terpolymer, however, initiates the formation of collagen nano-fibres in the microcapsule.…”

Section: Effect Of Collagen Methylationmentioning

confidence: 99%

See 2 more Smart Citations

Optimization of 3-D hepatocyte culture by controlling the physical and chemical properties of the extra-cellular matrices

Zhou³

et al. 2005

Biomaterials

Self Cite

View full text Add to dashboard Cite

Section: Imaging and Characterization Of Nano-fibres In Microcapsulementioning

confidence: 99%

Section: Imaging and Characterization Of Nano-fibres In Microcapsulementioning

confidence: 99%

Section: Effect Of Collagen Methylationmentioning

confidence: 99%

See 1 more Smart Citation

Optimization of 3-D hepatocyte culture by controlling the physical and chemical properties of the extra-cellular matrices

Zhou³

et al. 2005

Biomaterials

Self Cite

View full text Add to dashboard Cite

“…Indeed, several studies provided preliminary, qualitative evidence that neurite outgrowth and branching of PC12 cells was inhibited with decreasing substrate rigidity (Leach et al 2007). On the other hand, attachment, morphology, and functions of PC12 cells were superior in 3-D compared to 2-D culture conditions (Chia et al 2005). These observations deserve further careful quantitative studies, for which the present GFP-P12 cells might be well suited.…”

Section: Discussionmentioning

confidence: 63%

Quantitative Assessment of Neuronal Differentiation in Three-dimensional Collagen Gels Using Enhanced Green Fluorescence Protein Expressing PC12 Pheochromocytoma Cells

et al. 2008

View full text Add to dashboard Cite

There is a paucity of quantitative methods for evaluating the morphological differentiation of neuronal cells in a three-dimensional (3-D) system to assist in quality control of neural tissue engineering constructs for use in reparative medicine. Neuronal cells tend to aggregate in the 3-D scaffolds, hindering the application of two-dimensional (2-D) morphological methods to quantitate neuronal differentiation. To address this problem, we developed a stable transfectant green fluorescence protein (GFP)-PC12 neuronal cell model, in which the differentiation process in 3-D can be monitored with high sensitivity by fluorescence microscopy. Under 2-D conditions, the green cells showed collagen adherence, round morphology, proliferation properties, expression of the nerve growth factor (NGF) receptors TrkA and p75(NTR), stimulation of extracellular signal-regulated kinase phosphorylation by NGF and were able to differentiate in a dose-dependent manner upon NGF treatment, like wild-type (wt)-PC12 cells. When grown within 3-D collagen gels, upon NGF treatment, the GFP-PC12 cells differentiated, expressing long neurite outgrowths. We describe here a new validated method to measure NGF-induced differentiation in 3-D. Having properties similar to those of wt-PC12 and an ability to grow and differentiate in 3-D structures, these highly visualized GFP-expressing PC12 cells may serve as an ideal model for investigating various aspects of differentiation to serve in neural engineering.

show abstract

“…Indeed, there are experimental evidence that attachment, morphology, and functions of PC12 cells are superior in three-dimensional (3D) scaffolds compared with monolayer culture conditions. 7 Hence, there is a strong interest in both academia and industry to identify assays for neuronal cell growth in 3D tissue-mimicking environments, including promising examples of laminin-coated agarose gels, 8 self-assembling beta-sheet oligopeptide scaffolds, 9 and collagen, laminin, and fibronectin gels. 10,11 The general consensus of 3D in vitro cultivation studies, comparing different materials, is that neurons grown in collagen hydrogels show longer periods of survival and better neurite extension than other hydrogels.…”

Section: Introductionmentioning

confidence: 99%

Neuronal Networks on Nanocellulose Scaffolds

Jönsson

Brackmann

Puchades

et al. 2015

Tissue Engineering Part C: Methods

View full text Add to dashboard Cite

Proliferation, integration, and neurite extension of PC12 cells, a widely used culture model for cholinergic neurons, were studied in nanocellulose scaffolds biosynthesized by Gluconacetobacter xylinus to allow a threedimensional (3D) extension of neurites better mimicking neuronal networks in tissue. The interaction with control scaffolds was compared with cationized nanocellulose (trimethyl ammonium betahydroxy propyl [TMAHP] cellulose) to investigate the impact of surface charges on the cell interaction mechanisms. Furthermore, coatings with extracellular matrix proteins (collagen, fibronectin, and laminin) were investigated to determine the importance of integrin-mediated cell attachment. Cell proliferation was evaluated by a cellular proliferation assay, while cell integration and neurite propagation were studied by simultaneous label-free Coherent anti-Stokes Raman Scattering and second harmonic generation microscopy, providing 3D images of PC12 cells and arrangement of nanocellulose fibrils, respectively. Cell attachment and proliferation were enhanced by TMAHP modification, but not by protein coating. Protein coating instead promoted active interaction between the cells and the scaffold, hence lateral cell migration and integration. Irrespective of surface modification, deepest cell integration measured was one to two cell layers, whereas neurites have a capacity to integrate deeper than the cell bodies in the scaffold due to their fine dimensions and amoeba-like migration pattern. Neurites with lengths of >50 mm were observed, successfully connecting individual cells and cell clusters. In conclusion, TMAHP-modified nanocellulose scaffolds promote initial cellular scaffold adhesion, which combined with additional cell-scaffold treatments enables further formation of 3D neuronal networks.

show abstract

Engineering microenvironment for expansion of sensitive anchorage-dependent mammalian cells

Cited by 8 publications

References 43 publications

Optimization of 3-D hepatocyte culture by controlling the physical and chemical properties of the extra-cellular matrices

Optimization of 3-D hepatocyte culture by controlling the physical and chemical properties of the extra-cellular matrices

Quantitative Assessment of Neuronal Differentiation in Three-dimensional Collagen Gels Using Enhanced Green Fluorescence Protein Expressing PC12 Pheochromocytoma Cells

Neuronal Networks on Nanocellulose Scaffolds

Contact Info

Product

Resources

About