A Cellophane-Strip Technique for Culturing Tissue in Multipurpose Culture Chambers

Rose, George G.; Pomerat, C. M.; Shindler, Thomas O.; Trunnell, J. B.

doi:10.1083/jcb.4.6.761

Cited by 212 publications

(77 citation statements)

References 7 publications

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“…TCPS cultures were maintained with medium exchanges every 2 days but without subculture. The bioreactor design was based on the 'simultaneous-growthand-dialysis' method pioneered by Rose in the early 1960's [113,114] and avoided both subculture and periodic media exchanges, so that the pericellular environment was extremely stable within the bioreactor. Fig.…”

Section: Long-term Viability Of Hfob In Culturementioning

confidence: 99%

Influence of substratum surface chemistry/energy and topography on the human fetal osteoblastic cell line hFOB 1.19: Phenotypic and genotypic responses observed in vitro☆

et al. 2007

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Time-dependent phenotypic response of a model osteoblast cell line (hFOB 1.19, ATCC, CRL-11372) to substrata with varying surface chemistry and topography is reviewed within the context of extant cell-adhesion theory. Cell-attachment and proliferation kinetics are compared using morphology as a leading indicator of cell phenotype. Expression of (α 2 , α 3 , α 4 , α 5 , α v , β 1 and β 3 ) integrins, vinculin, as well as secretion of osteopontin and type I collagen supplement this visual assessment of hFOB growth. It is concluded that significant cell-adhesion events -contact, attachment, spreading, and proliferation -are similar on all surfaces, independent of substratum surface chemistry/energy. However, this sequence of events is significantly delayed and attenuated on hydrophobic (poorly water-wettable) surfaces exhibiting characteristically low-attachment efficiency and long induction periods before cells engage in an exponential-growth phase. Results suggest that a 'time-cell-substratum compatibility-superposition-principle' is at work wherein similar bioadhesive outcomes can be ultimately achieved on all surface types with varying hydrophilicity, but the time required to arrive at this outcome increases with decreasing cellsubstratum compatibility. Genomic and proteomic tools offer unprecedented opportunity to directly measure changes in the cellular machinery that lead to observed cell responses to different materials. But for the purpose of measuring structure-property relationships that can guide biomaterial development, genomic/proteomic tools should be applied early in the adhesion/spreading process before cells have an opportunity to significantly remodel the cell-substratum interface, effectively erasing cause-and-effect relationships between cell cell-substratum compatibility and substratum properties.Impact Statement-This review quantifies relationships among cell phenotype, substratum surface chemistry/energy, topography, and cell-substratum contact time for the model osteoblast cell

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Section: Long-term Viability Of Hfob In Culturementioning

confidence: 99%

Influence of substratum surface chemistry/energy and topography on the human fetal osteoblastic cell line hFOB 1.19: Phenotypic and genotypic responses observed in vitro☆

et al. 2007

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“…Microinjected cells were mounted in a Rose chamber (Rose et al, 1958) in recording media supplemented with 40 l Oxyrase (Oxyrase, Mansfield, OH) per ml of medium to reduce photobleaching. Injected cells were identified by the presence of RITC fluorescence.…”

Section: Time-lapse Microscopy and Image Acquisitionmentioning

confidence: 99%

Modulation of Microtubule Dynamics by Tau in Living Cells: Implications for Development and Neurodegeneration

Bunker

Wilson

Jordan

et al. 2004

MBoC

134

127

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The neural microtubule-associated protein tau binds to and stabilizes microtubules. Because of alternative mRNA splicing, tau is expressed with either 3 or 4 C-terminal repeats. Two observations indicate that differences between these tau isoforms are functionally important. First, the pattern of tau isoform expression is tightly regulated during development. Second, mutation-induced changes in tau RNA splicing cause neuronal cell death and dementia simply by altering the isoform expression ratio. To investigate whether 3-and 4-repeat tau differentially regulate microtubule behavior in cells, we microinjected physiological levels of these two isoforms into EGFP-tubulin-expressing cultured MCF7 cells and measured the effects on the dynamic instability behavior of individual microtubules by time-lapse microscopy. Both isoforms suppressed microtubule dynamics, though to different extents. Specifically, 4-repeat tau reduced the rate and extent of both growing and shortening events. In contrast, 3-repeat tau stabilized most dynamic parameters about threefold less potently than 4-repeat tau and had only a minimal ability to suppress shortening events. These differences provide a mechanistic rationale for the developmental shift in tau isoform expression and are consistent with a loss-of-function model in which abnormal tau isoform expression results in the inability to properly regulate microtubule dynamics, leading to neuronal cell death and dementia. INTRODUCTIONThe microtubule-associated protein tau promotes neuronal cell polarization and axonal outgrowth; it is also necessary for maintaining axonal morphology and axonal transport (Caceres and Kosik, 1990;Esmaeli-Azad et al., 1994;Trinczek et al., 1999;Stamer et al., 2002). Alternatively, abnormal tau is the major component of neurofibrillary tangles, hallmark pathological features of Alzheimer's disease and related dementias.Mechanistically, tau acts by binding to microtubules directly and regulating their growing, shortening, and treadmilling dynamics (Drechsel et al., 1992;Panda et al., 1995Panda et al., , 1999Trinczek et al., 1995). Because tight regulation of microtubule dynamics and microtubule behavior can be critical to cell viability (Jordan et al., 1996;Goncalves et al., 2001), fine regulation of tau activity may be equally critical.Tau activity is regulated in part by alternative mRNA splicing, which leads to the expression of two families of tau isoforms, those containing four C-terminal repeats and those with three such repeats ( Figure 1A; Lee et al., 1988;Himmler, 1989). In vitro studies have shown that 4-repeat tau binds to, assembles, and stabilizes microtubules more effectively than 3-repeat tau (Butner and Kirschner, 1991;Gustke et al., 1994;Trinczek et al., 1995;Goode et al., 2000;Panda et al., 2003). However, there are limited data assessing the abilities of 3-or 4-repeat tau to modulate dynamics in a complex cellular environment.The importance of functional differences between 3-and 4-repeat tau is highlighted by two observations. First, altho...

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“…Primary cultures of newt lung epithelial cells were grown on coverslips as described by Rose et al (1958) in chambers at room temperature (23°C) as described previously (Cassimeris et al, 1988). Cells were maintained in half-strength L-15 medium (pH 7.2) supplemented with 10% fetal bovine serum, 5 mM HEPES buffer, 100 U/ml penicillin, 0.1 mg/ml streptomycin, and 0.25 ,ug/ml amphotericin B. BSC-1 cells were cultured and grown on coverslips as described previously (Shelden and Wadsworth, 1993;.…”

Section: Cell Culturementioning

confidence: 99%

Nanomolar concentrations of nocodazole alter microtubule dynamic instability in vivo and in vitro.

Vasquez

Howell

Yvon

et al. 1997

MBoC

416

328

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Previous studies demonstrated that nanomolar concentrations of nocodazole can block cells in mitosis without net microtubule disassembly and resulted in the hypothesis that this block was due to a nocodazole-induced stabilization of microtubules. We tested this hypothesis by examining the effects of nanomolar concentrations of nocodazole on microtubule dynamic instability in interphase cells and in vitro with purified brain tubulin. Newt lung epithelial cell microtubules were visualized by video-enhanced differential interference contrast microscopy and cells were perfused with solutions of nocodazole ranging in concentration from 4 to 400 nM. Microtubules showed a loss of the two-state behavior typical of dynamic instability as evidenced by the addition of a third state where they exhibited little net change in length (a paused state). Nocodazole perfusion also resulted in slower elongation and shortening velocities, increased catastrophe, and an overall decrease in microtubule turnover. Experiments performed on BSC-1 cells that were microinjected with rhodamine-labeled tubulin, incubated in nocodazole for 1 h, and visualized by using low-light-level fluorescence microscopy showed similar results except that nocodazole-treated BSC-1 cells showed a decrease in catastrophe. To gain insight into possible mechanisms responsible for changes in dynamic instability, we examined the effects of 4 nM to 12 ,uM nocodazole on the assembly of purified tubulin from axoneme seeds. At both microtubule plus and minus ends, perfusion with nocodazole resulted in a dose-dependent decrease in elongation and shortening velocities, increase in pause duration and catastrophe frequency, and decrease in rescue frequency. These effects, which result in an overall decrease in microtubule turnover after nocodazole treatment, suggest that the mitotic block observed is due to a reduction in microtubule dynamic turnover. In addition, the in vitro results are similar to the effects of increasing concentrations of GDP-tubulin (TuD) subunits on microtubule assembly. Given that nocodazole increases tubulin GTPase activity, we propose that nocodazole acts by generating TuD subunits that then alter dynamic instability.

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A Cellophane-Strip Technique for Culturing Tissue in Multipurpose Culture Chambers

Cited by 212 publications

References 7 publications

Influence of substratum surface chemistry/energy and topography on the human fetal osteoblastic cell line hFOB 1.19: Phenotypic and genotypic responses observed in vitro☆

Influence of substratum surface chemistry/energy and topography on the human fetal osteoblastic cell line hFOB 1.19: Phenotypic and genotypic responses observed in vitro☆

Modulation of Microtubule Dynamics by Tau in Living Cells: Implications for Development and Neurodegeneration

Nanomolar concentrations of nocodazole alter microtubule dynamic instability in vivo and in vitro.

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