Spheroid Preparation from Hanging Drops: Characterization of a Model of Brain Tumor Invasion

Duca, Danny Del; Werbowetski, Tamra; Maestro, Rolando F. Del

doi:10.1023/b:neon.0000024220.07063.70

Cited by 222 publications

(134 citation statements)

References 18 publications

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“…To determine whether any of the genes induced by mutant K-Ras contributes to its transformation properties, we knocked down ETV1 and NT5E using siRNA pools in the same panel of 11 cell lines, and monitored proliferation in the second dimension on plastic and in the third dimension in hanging drop cultures (26). Knockdown relative to a nontargeting control is shown in supplemental Fig.…”

Section: Resultsmentioning

confidence: 99%

Identification of Mutant K-Ras-dependent Phenotypes Using a Panel of Isogenic Cell Lines

Vartanian¹,

Bentley²,

Brauer³

et al. 2013

Journal of Biological Chemistry

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Background: Many studies analyzing K-Ras function rely on overexpression. Results: Knock-out and knock-in of endogenous mutant K-ras have modest effects on downstream signaling but strong effects on gene expression and transformation. Conclusion: Genomic manipulation allows physiological determination of WT and mutant K-Ras consequences. Significance: Gene expression patterns can be used to monitor inhibition of mutant K-Ras.

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Section: Resultsmentioning

confidence: 99%

Identification of Mutant K-Ras-dependent Phenotypes Using a Panel of Isogenic Cell Lines

Vartanian¹,

Bentley²,

Brauer³

et al. 2013

Journal of Biological Chemistry

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“…25 Briefly, 30 mL drops of the culture medium containing cells (1 · 10 6 cells/mL) were applied onto the inside of the lid of a Petri dish that contained PBS to prevent drying out. After 24 h of incubation in an incubator, the spheroids formed in the drops were retrieved using a Pasteur pipette.…”

Section: Culture Of Hcbmsc Spheroidsmentioning

confidence: 99%

Transplantation of Cord Blood Mesenchymal Stem Cells as Spheroids Enhances Vascularization

Bhang

Lee

Shin

et al. 2012

Tissue Engineering Part A

174

183

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Despite promising results from the therapeutic use of stem cells for treating ischemic diseases, the poor survival of cells transplanted into ischemic regions is one of the major problems that undermine the efficacy of stem cell therapy. Cord blood mononuclear cells (CBMNCs) are an alternative source of mesenchymal stem cells (MSCs) without disadvantages, such as the painful and invasive harvesting procedure, of MSCs derived from bone marrow or adipose tissue. In the present study, we investigated whether the angiogenic efficacy of cord blood mesenchymal stem cells (CBMSCs) can be enhanced by grafting as spheroids in a mouse hindlimb ischemia model. Human CBMSC (hCBMSC) spheroids were prepared by using the hanging-drop method. Mouse hindlimb ischemia was induced by excising the femoral artery and its branches. After surgery, the animals were divided into no-treatment, dissociated hCBMSC, and spheroid hCBMSC groups (n=8 per group) and received corresponding hCBMSC treatments. After surgery, the ischemic hindlimbs were monitored for 4 weeks, and then, the ischemic hindlimb muscles were harvested for histological analysis. Apoptotic signaling, angiogenesis-related signal pathways, and blood vessel formation were investigated in vitro and/or in vivo. The transplantation of hCBMSCs as spheroids into mouse ischemic hindlimbs significantly improved the survival of the transplanted cells by suppressing apoptotic signaling while activating antiapoptotic signaling. Furthermore, the transplantation of hCBMSCs as spheroids significantly increased the number of microvessels and smooth muscle α-actin-positive vessels in the ischemic limbs of mice, and attenuated limb loss and necrosis. Human CBMNC can be considered an alternative source of MSC, and spheroid-based hCBMSC delivery can be considered a simple and effective strategy for enhancing the therapeutic efficacy of hCBMSCs.

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“…Many compositions of the matrix have been studied. One of the widely used constituents is the collagen [225][226][227], eventually mixed with other molecules, such as agarose [228], the ECM proteins tenascin [229] or chondroitin sulfate [230]. Among the ECM components, an important role has been demonstrated for HA, able to regulate the glioma cell phenotype [231] or to increase stem-like properties of glioblastoma cells [232].…”

Section: Tumoral Cns Cultures On Scaffoldsmentioning

confidence: 99%

“…The widely used 3D transwell or spheroid invasion/motility assays in soft matrices, like collagen, Matrigel TM , fibronectin or agar are extensively applied in brain tumor research [225,226,[240][241][242][243][244]. Moreover, several other matrix compositions have been tested with the aim of better reproducing the in vivo microenvironment.…”

Section: Models For Testing Motility/invasion Of Cns Tumorsmentioning

confidence: 99%

In Vitro Modeling of Tissue-Specific 3D Microenvironments and Possibile Application to Pediatric Cancer Research

Pizzimenti¹

2014

J. Pediatr. Oncol.

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A large body of evidence indicates that three dimensional (3D) cancer models are superior to two-dimensional (2D) ones in better representing the in vivo phenomena. Indeed, 3D models allow recapitulating in vitro the in vivo features observed in solid tumors (e.g. cell polarity, cell-cell/cell-matrix interactions, biochemical/metabolic gradients, anchorage-independent growth and hypoxia). Moreover, it is well established that the microenvironment plays a fundamental role in regulating tumor development and behavior, including drug resistance. Thus, innovative models able to mimic this complexity represent attractive tools in cancer research. In this review article, we provide a comprehensive review of the application of 3D culture systems in pediatrics' cancer research. In particular, 3D in vitro/ex vivo models of the most common pediatric tumors, such as leukemias, lymphomas and malignancies of the nervous system, will be considered.

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Spheroid Preparation from Hanging Drops: Characterization of a Model of Brain Tumor Invasion

Cited by 222 publications

References 18 publications

Identification of Mutant K-Ras-dependent Phenotypes Using a Panel of Isogenic Cell Lines

Identification of Mutant K-Ras-dependent Phenotypes Using a Panel of Isogenic Cell Lines

Transplantation of Cord Blood Mesenchymal Stem Cells as Spheroids Enhances Vascularization

In Vitro Modeling of Tissue-Specific 3D Microenvironments and Possibile Application to Pediatric Cancer Research

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