Predicting Aggregation Kinetics of DU 145 Prostate Cancer Cells in Liquid-Overlay Culture

O’Connor, Kim C.; Venczel, Mark

doi:10.1007/s10529-005-2725-6

Cited by 8 publications

(9 citation statements)

References 15 publications

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“…The formation of detached cells was maintained in culture under suspended conditions, and individual aggregations for every phase were randomly selected, analyzed and measured in triplicate experiments. The longest diameters of cells with irregular aggregations were determined by NIS-Elements F2.30 software according to the procedure described by Conner [6]. Trypan blue exclusion assay was used to evaluate the viability of cells at different time point, and the morphological changes were recorded by an Optronics digital camera, Nikon 8700.…”

Section: Introductionmentioning

confidence: 99%

Aggregation formation mediated anoikis resistance of BEL7402 hepatoma cells.

Zhang

Han²,

Cao³

et al. 2008

Folia Histochem Cytobiol.

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Materials and methodCell culture and reagents. The human hepatoma cell line BEL7402 was kept in institute of Immunology, School of Medicine, Shandong University. Hepatoma cells were maintained in RMPI1640 (Gibco, Grand Island, N.Y.), supplemented with 10% fetal bovine serum (FuMeng, Shanghai). Poly-HEMA (Sigma) was prepared by dissolving in 95% ethanol to a concentration of 12 mg/ml, coated 96-well plate with 0.1 ml/cm 2 of surface and allowed to air dry in a sterile environment. Attached and detached BEL7402 cells were generated by sequential cycles of culture on untreated (adhered) and poly-HEMA treated cell culture wells (suspended). Both the detached and attached cells were plat- Abstract: Anoikis resistance is the prerequisite of cancer cells metastasis. Elucidation of the mechanism of anoikis resistance remains a significant challenge. We reported here a model to mimic anoikis resistant process of hepatoma cells in vitro. Experimental results indicated cell to cell aggregation could mediate anoikis resistance of BEL7402 hepatoma cells. Further investigation of these aggregations indicated the biological properties changed greatly after the hepatoma cells lost their anchorage. Aggregation forming process could be separated into three distinct phases according to their biological characteristics, comprising of premature phase, mature phase and postmature phase. Mature phase aggregations have the premium state of cell viability and may mimic the metastatic cells in the circulating system. Biological properties of these three phases aggregations were studied in details including morphological alteration, cell viability and microarray expression profiles. It indicated there was a great upregulation of adhesion molecules during the process of aggregation formation and the cell to cell contact in the aggregation may be mediated independent of calcium involved adhesion pathway. This model might shed light on the anoikis resistance mechanism of hepatoma cells and help to develop new therapies that may target the anoikis resistant hepatoma cells in the metastasis process.

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Section: Introductionmentioning

confidence: 99%

Aggregation formation mediated anoikis resistance of BEL7402 hepatoma cells.

Zhang

Han²,

Cao³

et al. 2008

Folia Histochem Cytobiol.

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“…In this study, cells were cultured with a liquid overlay technique [12,13] . After the for mation of multicellular spheroids, morphology was analyzed by phase contrast m i c r o s c o p y, s c a n n i n g e l e c t r o n m i c r o s c o p y a n d transmission electron microscopy.…”

Section: Introductionmentioning

confidence: 99%

Chinese medicinal compound delisheng has satisfactory anti-tumor activity, and is associated with up-regulation of endostatin in human hepatocellular carcinoma cell line HepG2 in three-dimensional culture

Cui

Kang²,

Tian³

et al. 2007

WJG

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AIM:To investigate the multicellular resistance of human hepatocellular carcinoma HepG2 cells in threedimensional culture to delisheng, 5-fluorouracil and adriamycin, and the possible molecular mechanisms of delisheng. METHODS:Human hepatocellular carcinoma HepG2 cells were cultured with a liquid overlay technique. After the formation of multicellular spheroids, morphology was analyzed by phase contrast microscopy, scanning electron microscopy and transmission electron microscopy. Sensitivity of HepG2 cells to delisheng, 5-fluorouracil and adriamycin was investigated by MTT assay in multicelluar spheroids and monolayers. Vascular endothelial growth factor (VEGF) and endostatin expression were analyzed in multicellular spheroids treated with delisheng, 5-fluorouracil, adriamycin and negative control PBS, with immunohistochemical staining. RESULTS:Multicellular spheroids exhibited structural c h a ra c t e r i s t i c s s o m e w h a t d i f f e r e n t t o t h o s e i n monolayers. The cells in three-dimensional cell culture turned out to be less sensitive to delisheng, 5-fluorouracil and adriamycin than the cells cultured in monolayer. This showed that delisheng had a satisfactory cells inhibition ratio compared to 5-fluorouracil and adriamycin. Immunohistochemical staining showed that VEGF and endostatin expression was positive during growth as multicellular spheroids, and endostatin expression in spheroids with treatment of delisheng was higher than that with 5-fluorouracil, adriamycin and PBS (139.35 ± 7.83, 159.23 ± 10.34, 162.83 ± 3.47 and 148.48 ± 11.06, P < 0.05). CONCLUSION:Chinese medicine compound delisheng has satisfactory anti-tumor activity in HepG2 cells in three-dimensional culture, and the effects are associated with up-regulation of endostatin.

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“…In the liquid overlay technique, tumor cells are placed in a gel composed of agarose or another biopolymer. The gel is fully submerged in a growth medium with nutrients that reach tumor cells through diffusion in the gel . Cell spinning methods can result in larger spheroids (< 2 mm) than other existing techniques can produce, but they require expensive equipment to prevent excessive shear to the cells …”

mentioning

confidence: 99%

“…The pellets were then transferred to the PDMS wells (one hanging drop culture per one PDMS well) to further grow into larger aggregates. The centrifugation to pellet tumor cells has previously been used to generate initial tumor spheroids …”

mentioning

confidence: 99%

“…The gel is fully submerged in a growth medium with nutrients that reach tumor cells through diffusion in the gel. 11 Cell spinning methods can result in larger spheroids (< 2 mm) than other existing techniques can produce, but they require expensive equipment to prevent excessive shear to the cells. 12 Soft lithographic procedures have been utilized to create synthetic scaffolds for spheroid production 13,14 or hemi-Correspondence concerning this article should be addressed to Damir B. Khismatullin at damir@tulane.edu spherical "rifts" on microchannel walls.…”

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confidence: 99%

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PDMS well platform for culturing millimeter‐size tumor spheroids

Ratnayaka

Hillburn²,

Forouzan³

et al. 2013

Biotechnology Progress

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Multicellular tumor spheroids are widely used as in vitro models for testing of anticancer drugs. The advantage of this approach is that it can predict the outcome of a drug treatment on human cancer cells in their natural three-dimensional environment without putting actual patients at risk. Several methods were utilized in the past to grow submillimeter-size tumor spheroids. However, these small models are not very useful for preclinical studies of tumor ablation where the goal is the complete destruction of tumors that can reach several centimeters in diameter in the human body. Here, we propose a PDMS well method for large tumor spheroid culture. Our experiments with HepG2 hepatic cancer cells show that three-dimensional aggregates of tumor cells with a volume as large as 44 mm(3) can be grown in cylindrical PDMS wells after the initial culture of tumor cells by the hanging drop method. This is a 350 times more than the maximum volume of tumor spheroids formed inside hanging drops (0.125 mm(3)).

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Predicting Aggregation Kinetics of DU 145 Prostate Cancer Cells in Liquid-Overlay Culture

Cited by 8 publications

References 15 publications

Aggregation formation mediated anoikis resistance of BEL7402 hepatoma cells.

Aggregation formation mediated anoikis resistance of BEL7402 hepatoma cells.

Chinese medicinal compound delisheng has satisfactory anti-tumor activity, and is associated with up-regulation of endostatin in human hepatocellular carcinoma cell line HepG2 in three-dimensional culture

PDMS well platform for culturing millimeter‐size tumor spheroids

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