Processing cellular spheroids for histological examination

Olsen, Timothy R.; Mattix, Brandon; Casco, Megan; Herbst, Austin; Williams, Colby.; Tarasidis, Anna.; Evans, Grace; Jenkins, Linda; McMahan, C.; Simionescu, Dan T.; Visconti, Richard P.; Alexis, Frank

doi:10.1179/2046023614y.0000000047

Cited by 16 publications

(19 citation statements)

References 15 publications

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“…All samples were processed and sectioned via paraffin sectioning techniques developed in our lab [23]. Spheroids were fixed, processed (5 μm sections) and stained using either hemotoxylin and eosin (H&E) or Masson’s Trichrome stain.…”

Section: Methodsmentioning

confidence: 99%

Manipulation of cellular spheroid composition and the effects on vascular tissue fusion

et al. 2015

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Cellular spheroids were investigated as tissue-engineered building blocks that can be fused to form functional tissue constructs. While spheroids can be assembled using passive contacts for the fusion of complex tissues, physical forces can be used to promote active contacts to improve tissue homogeneity and accelerate tissue fusion. Understanding the mechanisms affecting the fusion of spheroids is critical to fabricating tissues. Here, manipulation of the spheroid composition was used to accelerate the fusion process mediated by magnetic forces. The Janus structure of magnetic cellular spheroids spatially controls iron oxide magnetic nanoparticles (MNPs) to form two distinct domains: cells and extracellular MNPs. Studies were performed to evaluate the influence of extracellular matrix (ECM) content and cell number on the fusion of Janus magnetic cellular spheroids (JMCSs). Results showed that the integration of iron oxide MNPs into spheroids increased the production of collagen over time when compared to spheroids without MNPs. The results also showed that ring tissues composed of JMCSs with high ECM concentrations and high cell numbers fused together, but exhibited less contraction when compared to their lower concentration counterparts. Results from spheroid fusion in capillary tubes showed that low ECM concentrations and high cell numbers experienced more fusion and cellular intermixing over time when compared to their higher counterparts. These findings indicate that cell–cell and cell–matrix interactions play an important role in regulating fusion, and this understanding sets the rationale of spheroid composition to fabricate larger and more complex tissue-engineered constructs.

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Section: Methodsmentioning

confidence: 99%

Manipulation of cellular spheroid composition and the effects on vascular tissue fusion

et al. 2015

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“…CyGEL restricts spheroid movement without compromising the viability, morphology, and protein expression in spheroids (Robertson et al , 2010). Additionally, the small size and fragile nature of spheroids require special fixing and sectioning techniques (Olsen et al , 2014). High quality confocal images are obtained by culturing spheroids on clear glass bottom plates and using wet immersion objective (Roux et al , 2008).…”

Section: Tools For Characterization Of Mctsmentioning

confidence: 99%

Three-dimensional culture systems in cancer research: Focus on tumor spheroid model

Nath

Devi

2016

Pharmacology & Therapeutics

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667

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Cancer cells propagated in three-dimensional (3D) culture systems exhibit physiologically relevant cell-cell and cell-matrix interactions, gene expression and signaling pathway profiles, heterogeneity and structural complexity that reflect in vivo tumors. In recent years, development of various 3D models have improved the study of host-tumor interaction and use of high-throughput screening platforms for anti-cancer drug discovery and development. This review attempts to summarize the various 3D culture systems, with an emphasis on the most well characterized and widely applied model - multicellular tumor spheroids. This review also highlights the various techniques to generate tumor spheroids, methods to characterize them, and its applicability in cancer research.

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“…After overnight fixation, the Z-Fix was removed and the tissue samples were dehydrated using a series of ethanol and xylene incubations (Table 1) [14]. Immediately prior to the first ethanol incubation, the tissue tube was removed from the silicone mandrel.…”

Section: Tissue Fixation Processing Embedding and Sectioningmentioning

confidence: 99%

“…Citation:Olsen TR, Casco M, Mattix B, Williams C, Herbst A, et al, (2014) Preparing and Processing Complex Tissue Constructs Composed of Cellular Spheroids for Histological Examination. J Cytol Tissue Biol 1: 002.…”

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confidence: 99%

Preparing and Processing Complex Tissue Constructs Composed of Cellular Spheroids for Histological Examination

Alexis¹

2014

CTB

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Using cellular spheroids to build tissues and organs is a rapidly emerging technology that will have great impact on the biomedical industry. As tissues are fabricated in vitro, it will be critical to histologically examine tissue composition, function and organization for comparison with native tissues. Conventional automated histologically methods are not recommended for fixation and processing of tissues composed of spheroids due to their delicate nature and the need to preserve tissue morphology. Given this, there is a need to develop a manual method to prepare and process tissues composed of cellular spheroids for histological sectioning and examination. Here, we report a manual processing technique developed for preparing larger tissues composed of spheroids for histological examination based on our lab's work using magnetic forces to pattern and assemble tissues composed of Janus Magnetic Cellular Spheroids for tissue engineering applications. The results presented here contribute to understanding the organization of cells, studying extracellular matrix content, analyzing phenotypic expression and localizing iron oxide magnetic nanoparticles in tissues composed of Janus Magnetic Cellular Spheroids.

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Processing cellular spheroids for histological examination

Cited by 16 publications

References 15 publications

Manipulation of cellular spheroid composition and the effects on vascular tissue fusion

Manipulation of cellular spheroid composition and the effects on vascular tissue fusion

Three-dimensional culture systems in cancer research: Focus on tumor spheroid model

Preparing and Processing Complex Tissue Constructs Composed of Cellular Spheroids for Histological Examination

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