scholarly journals Rapid spheroid assays in a 3-dimensional cell culture chip

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Jia Lin Teh ◽  
Siti Fairus Abdul Rahman ◽  
Gregory Domnic ◽  
Lengishwarra Satiyasilan ◽  
Nelson Jeng Yeou Chear ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Biology ◽  
Drug Sensitivity ◽  
3D Cell Culture ◽  
Collagen Matrix ◽  
High Volume ◽  
Collagen I ◽  
Bovine Collagen ◽  
High Concentrations ◽  
Drug Compound

Abstract Objective The spheroid model provides a physiological platform to study cancer cell biology and drug sensitivity. Usage of bovine collagen I for spheroid assays is costly especially when experiments are conducted in 24-well plates, as high volume of bovine collagen I is needed. The aim of the study was to downsize spheroid assays to a microfluidic 3D cell culture chip and compare the growth, invasion and response to drug/compound of spheroids embedded in the 3D chip to spheroids embedded in 24-well plates. Results Spheroids generated from nasopharyngeal carcinoma cell line HK-1 continuously grew and invaded into collagen matrix in a 24-well plate. Similar observations were noticed with spheroids embedded in the 3D chip. Large spheroids in both 24-well plate and the 3D chip disintegrated and invaded into the collagen matrix. Preliminary drug sensitivity assays showed that the growth and invasion of spheroids were inhibited when spheroids were treated with combination of cisplatin and paynantheine at high concentrations, in a 24-well plate. Comparable findings were obtained when spheroids were treated with the same drug combination in the 3D chip. Moving forward, spheroid assays could be performed in the 3D chip in a more high-throughput manner with minimal time and cost.

scholarly journals Rapid Spheroid Assays in a 3-Dimensional Cell Culture Chip

2020 ◽  
Author(s):  
Jia Lin Teh ◽  
Siti Fairus Abdul Rahman ◽  
Gregory Domnic ◽  
Lengishwarra Satiyasilan ◽  
Nelson Jeng Yeou Chear ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Biology ◽  
Drug Sensitivity ◽  
3D Cell Culture ◽  
Collagen Matrix ◽  
High Volume ◽  
Collagen I ◽  
Bovine Collagen ◽  
High Concentrations ◽  
Drug Compound

Abstract Objective: The spheroid model provides a physiological platform to study cancer cell biology and drug sensitivity. Usage of bovine collagen I for spheroid assays is costly especially when experiments are conducted in 24-well plates, as high volume of bovine collagen I is needed. The aim of the study was to downsize spheroid assays to a microfluidic 3D cell culture chip and compare the growth, invasion and response to drug/compound of spheroids embedded in the 3D chip to spheroids embedded in 24-well plates.Results: Spheroids generated from nasopharyngeal carcinoma cell line HK-1 continuously grew and invaded into collagen matrix in a 24-well plate. Similar observations were noticed with spheroids embedded in the 3D chip. Large spheroids in both 24-well plate and the 3D chip disintegrated and invaded into the collagen matrix. Preliminary drug sensitivity assays showed that the growth and invasion of spheroids were inhibited when spheroids were treated with combination of cisplatin and paynantheine at high concentrations, in a 24-well plate. Comparable findings were obtained when spheroids were treated with the same drug combination in the 3D chip. Moving forward, spheroid assays could be performed in the 3D chip in a more high-throughput manner with minimal time and cost.

Ratiometric Nanoviscometers: Applications for Measuring Cellular Physical Properties in 3D Cultures

2020 ◽  
Vol 25 (3) ◽  
pp. 234-246
Author(s):  
Charles McRae White ◽  
Mark A. Haidekker ◽  
William S. Kisaalita
Keyword(s):  
Cell Culture ◽  
Cell Cultures ◽  
Cell Biology ◽  
Low Cost ◽  
3D Cell Culture ◽  
Biomechanical Properties ◽  
Practical Applications ◽  
3D Cell Cultures

New insights into the biomechanical properties of cells are revealing the importance of these properties and how they relate to underlying molecular, architectural, and behavioral changes associated with cell state and disease processes. However, the current understanding of how these in vitro biomechanical properties are associated with in vivo processes has been developed based on the traditional monolayer (two-dimensional [2D]) cell culture, which traditionally has not translated well to the three-dimensional (3D) cell culture and in vivo function. Many gold standard methods and tools used to observe the biomechanical properties of 2D cell cultures cannot be used with 3D cell cultures. Fluorescent molecules can respond to external factors almost instantaneously and require relatively low-cost instrumentation. In this review, we provide the background on fluorescent molecular rotors, which are attractive tools due to the relationship of their emission quantum yield with environmental microviscosity. We make the case for their use in both 2D and 3D cell cultures and speculate on their fundamental and practical applications in cell biology.

scholarly journals PDX-Derived Ewing’s Sarcoma Cells Retain High Viability and Disease Phenotype in Alginate Encapsulated Spheroid Cultures

Cancers ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 879
Author(s):  
Giacomo Domenici ◽  
Rodrigo Eduardo ◽  
Helena Castillo-Ecija ◽  
Gorka Orive ◽  
Ángel Montero Carcaboso ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Lines ◽  
Drug Sensitivity ◽  
Ewing’S Sarcoma ◽  
Ewing's Sarcoma ◽  
3D Cell Culture ◽  
Suitable Model ◽  
Es Cell ◽  
3D Cultures ◽  
Novel Drugs

Ewing’s Sarcoma (ES) is the second most frequent malignant bone tumour in children and young adults and currently only untargeted chemotherapeutic approaches and surgery are available as treatment, although clinical trials are on-going for recently developed ES-targeted therapies. To study ES pathobiology and develop novel drugs, established cell lines and patient-derived xenografts (PDX) are the most employed experimental models. Nevertheless, the establishment of ES cell lines is difficult and the extensive use of PDX raises economic/ethical concerns. There is a growing consensus regarding the use of 3D cell culture to recapitulate physiological and pathophysiological features of human tissues, including drug sensitivity. Herein, we implemented a 3D cell culture methodology based on encapsulation of PDX-derived ES cell spheroids in alginate and maintenance in agitation-based culture systems. Under these conditions, ES cells displayed high proliferative and metabolic activity, while retaining the typical EWSR1-FLI1 chromosomal translocation. Importantly, 3D cultures presented reduced mouse PDX cell contamination compared to 2D cultures. Finally, we show that these 3D cultures can be employed in drug sensitivity assays, with results similar to those reported for the PDX of origin. In conclusion, this novel 3D cell culture method involving ES-PDX-derived cells is a suitable model to study ES pathobiology and can assist in the development of novel drugs against this disease, complementing PDX studies.

Rapid screening for individualized chemotherapy optimization of colorectal cancer: a novel conditional reprogramming technology-based functional diagnostic assay.

2020 ◽  
Author(s):  
YingJie Li ◽  
Dagang Guo ◽  
Yihong Zhang ◽  
Lin Wang ◽  
Tingting Sun ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Culture ◽  
Cell Biology ◽  
Drug Sensitivity ◽  
Primary Cell Culture ◽  
Primary Cell ◽  
Rapid Screening ◽  
Primary Colorectal Tumor ◽  
New Generation

Abstract In vitro patient tumor models such as patient-derived organoids (PDO) and conditionally reprogrammed (CR) cell culture are important for translational research and pre-clinical drug testing.In this study we present a personalized drug sensitivity test for late stage, potentially operable colorectal cancer (CRC) using patient-derived primary cell culture based on a new generation CR technology. We explored the clinical feasibility of using CR-based primary cell culture system to guide CRC chemotherapy, and established the correlation between in vitro drug sensitivity and patient clinical response.Our novel CR platform (termed i-CR) can be used to propagate primary colorectal tumor cells that represent individual patient tumors effectively by keeping the clonal heterogeneity and comparable drug responses.Therefore, our platform can be used to test and optimize therapeutic regimens pre-clinically, study cancer cell biology, and model tumor re-emergence to identify new targeted therapeutics from an effective personalized medicine standpoint.

Predicting drug sensitivity by 3D cell culture models

2015 ◽  
Vol 8 (1) ◽  
pp. 77-80 ◽  
Author(s):  
Arno Amann ◽  
Gabriele Gamerith ◽  
Julia M. Huber ◽  
Marit Zwierzina ◽  
Wolfgang Hilbe ◽  
...  
Keyword(s):  
Cell Culture ◽  
Drug Sensitivity ◽  
3D Cell Culture ◽  
Culture Models ◽  
Cell Culture Models

A Novel Suspended Hydrogel Membrane Platform for Cell Culture

2015 ◽  
Vol 6 (2) ◽  
Author(s):  
Yong X. Chen ◽  
Shihao Yang ◽  
Jiahan Yan ◽  
Ming-Han Hsieh ◽  
Lingyan Weng ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Biology ◽  
3D Cell Culture ◽  
Cell Encapsulation ◽  
Cell Types ◽  
Ease Of Use ◽  
In Vitro Assays ◽  
Hydrogel Membrane

Current cell-culture is largely performed on synthetic two-dimensional (2D) petri dishes or permeable supports such as Boyden chambers, mostly because of their ease of use and established protocols. It is generally accepted that modern cell biology research requires new physiologically relevant three-dimensional (3D) cell culture platform to mimic in vivo cell responses. To that end, we report the design and development of a suspended hydrogel membrane (ShyM) platform using gelatin methacrylate (GelMA) hydrogel. ShyM thickness (0.25–1 mm) and mechanical properties (10–70 kPa) can be varied by controlling the size of the supporting grid and concentration of GelMA prepolymer, respectively. GelMA ShyMs, with dual media exposure, were found to be compatible with both the cell-seeding and the cell-encapsulation approach as tested using murine 10T1/2 cells and demonstrated higher cellular spreading and proliferation as compared to flat GelMA unsuspended control. The utility of ShyM was also demonstrated using a case-study of invasion of cancer cells. ShyMs, similar to Boyden chambers, are compatible with standard well-plates designs and can be printed using commonly available 3D printers. In the future, ShyM can be potentially extended to variety of photosensitive hydrogels and cell types, to develop new in vitro assays to investigate complex cell–cell and cell–extracellular matrix (ECM) interactions.

Commercializing Electrospun Scaffolds For Pluripotent Stem Cell-Based Tissue Engineering Applications

2017 ◽  
Vol 2 (1) ◽  
pp. 62-72 ◽  
Author(s):  
Nima Khadem Mohtaram ◽  
Vahid Karamzadeh ◽  
Yousef Shafieyan ◽  
Stephanie M. Willerth
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Cell Culture ◽  
Stem Cell ◽  
Cell Biology ◽  
3D Cell Culture ◽  
Bioactive Scaffolds ◽  
Electrospun Scaffolds ◽  
Current State ◽  
Start Up

Abstract Tissue engineering, the process of combining bioactive scaffolds often with cells to produce replacements for damaged organs, represents an enormous market opportunity. This review critically evaluates the commercialization potential of electrospun scaffolds for applications in stem cell biology, including tissue engineering. First, it provides an overview of pluripotent stem cells (PSCs) and their defining properties, pluripotency and the ability to self-renew. These cells serve as an important tool for engineering tissues, including for clinical applications. Next, we review the technique of electrospinning and its promise for fabricating cell culture substrates and scaffolds for directing tissue formation from stem cells and compare these scaffolds to existing technologies, such as hydrogels. We address the associated market for electrospun scaffolds for PSCs and its potential for growth along with highlighting the importance of 3D cell culture substrates for PSCs by analyzing the net capital invested in this market and the associated growth rate. This review finishes by detailing the current state of commercializing electrospun scaffolds along with pathways for translating these scaffolds from research laboratories into successful start-up companies and the associated challenges with this process.

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