Growth advantage (“clonal dominance”) of metastatically competent tumor cell variants expressed under selective two- or three-dimensional tissue culture conditions

1993 ◽  
Vol 29 (9) ◽  
pp. 742-748 ◽  
Author(s):  
Janusz W. Rak ◽  
Robert S. Kerbel
Keyword(s):  
Tissue Culture ◽  
Tumor Cell ◽  
Three Dimensional ◽  
Culture Conditions

scholarly journals Enrichment of Melanoma Stem-Like Cells via Sphere Assays

2021 ◽  
pp. 185-199
Author(s):  
Nabanita Mukherjee ◽  
Karoline A. Lambert ◽  
David A. Norris ◽  
Yiqun G. Shellman
Keyword(s):  
Cell Culture ◽  
Tissue Culture ◽  
Stem Cell ◽  
Three Dimensional ◽  
Culture Conditions ◽  
Low Density ◽  
In Vitro Techniques ◽  
Sphere Culture

AbstractSphere assays are widely used in vitro techniques to enrich and evaluate the stem-like cell behavior of both normal and cancer cells. Utilizing three-dimensional in vitro sphere culture conditions provide a better representation of tumor growth in vivo than the more common monolayer cultures. We describe how to perform primary and secondary sphere assays, used for the enrichment and self-renewability studies of melanoma/melanocyte stem-like cells. Spheres are generated by growing melanoma cells at low density in nonadherent conditions with stem cell media. We provide protocols for preparing inexpensive and versatile polyHEMA-coated plates, setting up primary and secondary sphere assays in almost any tissue culture format and quantification methods using standard inverted microscopy. Our protocol is easily adaptable to laboratories with basic cell culture capabilities, without the need for expensive fluidic instruments.

scholarly journals Barley somatic embryogenesis-an attempt to modify variation induced in tissue culture

2021 ◽  
Vol 28 (1) ◽  
Author(s):  
Renata Orłowska
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Taguchi Method ◽  
Culture Conditions ◽  
Dna Demethylation ◽  
Silver Ion ◽  
Donor Plant ◽  
In Vitro Tissue Culture ◽  
Induced Variation

Abstract Background Somatic embryogenesis is a phenomenon carried out in an environment that generates abiotic stress. Thus, regenerants may differ from the source of explants at the morphological, genetic, and epigenetic levels. The DNA changes may be the outcome of induction media ingredients (i.e., copper and silver ions) and their concentrations and time of in vitro cultures. Results This study optimised the level of copper and silver ion concentration in culture media parallel with the induction medium longevity step towards obtaining barley regenerants via somatic embryogenesis with a minimum or maximum level of tissue culture-induced differences between the donor plant and its regenerants. The optimisation process is based on tissue culture-induced variation evaluated via the metAFLP approach for regenerants derived under varying in vitro tissue culture conditions and exploited by the Taguchi method. In the optimisation and verification experiments, various copper and silver ion concentrations and the different number of days differentiated the tested trials concerning the tissue culture-induced variation level, DNA demethylation, and de novo methylation, including symmetric (CG, CHG) and asymmetric (CHH) DNA sequence contexts. Verification of optimised conditions towards obtaining regenerants with minimum and maximum variability compared to donor plants proved useful. The main changes that discriminate optimised conditions belonged to DNA demethylation events with particular stress on CHG context. Conclusions The combination of tissue culture-induced variation evaluated for eight experimental trials and implementation of the Taguchi method allowed the optimisation of the in vitro tissue culture conditions towards the minimum and maximum differences between a source of tissue explants (donor plant) and its regenerants from somatic embryos. The tissue culture-induced variation characteristic is mostly affected by demethylation with preferences towards CHG sequence context.

scholarly journals Effects of Natural Antioxidants on Phospholipid and Ceramide Profiles of 3D-Cultured Skin Fibroblasts Exposed to UVA or UVB Radiation

Antioxidants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 578
Author(s):  
Agnieszka Gęgotek ◽  
Wojciech Łuczaj ◽  
Elżbieta Skrzydlewska
Keyword(s):  
Ascorbic Acid ◽  
High Performance ◽  
Three Dimensional ◽  
Natural Antioxidants ◽  
Phospholipid Metabolism ◽  
Culture Conditions ◽  
Membrane Properties ◽  
Dimensional System ◽  
Uvb Radiation ◽  
Liquid Chromatograph

Ultraviolet (UV) radiation is one of the primary factors responsible for disturbances in human skin cells phospholipid metabolism. Natural compounds that are commonly used to protect skin, due to their lipophilic or hydrophilic nature, show only a narrow range of cytoprotective activity, which prompts research on their combined application. Therefore, the aim of this study was to examine the effect of ascorbic acid and rutin on the phospholipid and ceramide profiles in UV-irradiated fibroblasts cultured in a three-dimensional system that approximates the culture conditions to the dermis. An ultra-high-performance liquid chromatograph coupled with a quadrupole time-of-flight mass spectrometer was used for phospholipid and ceramide profiling. As a result of UVA and UVB cells irradiation, upregulation of phosphatidylcholines, ceramides, and downregulation of sphingomyelins were observed, while treatment with ascorbic acid and rutin of UVA/UVB-irradiated fibroblast promoted these changes to provide cells a stronger response to stress. Moreover, an upregulation of phosphatidylserines in cells exposed to UVB and treated with both antioxidants suggests the stimulation of UV-damaged cells apoptosis. Our findings provide new insight into action of rutin and ascorbic acid on regulation of phospholipid metabolism, which improves dermis fibroblast membrane properties.

Poly-(lactic acid) and fibrin bioactive cellularized scaffold for use in bone regenerative medicine: Proof of concept

2021 ◽  
pp. 088391152199640
Author(s):  
Renata Aquino de Carvalho ◽  
Valmir Vieira Rocha Júnior ◽  
Antonio José Felix Carvalho ◽  
Heloisa Sobreiro Selistre de Araújo ◽  
Mônica Rosas Costa Iemma ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Regenerative Medicine ◽  
Platelet Rich Plasma ◽  
Three Dimensional ◽  
Culture Conditions ◽  
Infrapatellar Fat Pad ◽  
Dimensional Structure ◽  
Poly Lactic Acid ◽  
Proof Of Concept ◽  
Critical Bone Defects

Bone regenerative medicine (BRM) aims to overcome the limitations of conventional treatments for critical bone defects by developing therapeutic strategies, based on temporary bioactive substitutes, capable of stimulating, sustaining, and guiding tissue regeneration. The aim of this study was to validate the “proof of concept” of a cellularized bioactive scaffold and establish its potential for use in BRM. For this purpose, three-dimensional scaffolds of poly-(lactic acid) (PLA), produced by the additive manufacturing technique, were incorporated into a human platelet-rich plasma (PRP-h) fibrin matrix containing human infrapatellar fat pad mesenchymal stem cells (hIFPMSC). The scaffolds (PLA/finbrin-bioactive) were kept under ideal culture conditions in a medium free from fetal bovine serum and analyzed at 5 and 10 days by Scanning Electron Microscopy (SEM), Fourrier Transform Infrared (FTIR), Circular Dichroism and fluorescence microscopy. The results demonstrated the feasibility of obtaining a rigid, cytocompatible, and cellularized three-dimensional structure. In addition, PRP platelets and leukocytes were able to provide a bioactive environment capable of maintaining the viability of hIFPMSC into scaffolds. The results validate the concept of a customizable, bioactive, cellularized, and non-immunogenic strategy for application in BRM.

Enhanced growth and myogenic differentiation of spheroid-derived C2C12 cells

2021 ◽  
Author(s):  
Guang-Zhen Jin
Keyword(s):  
Myogenic Differentiation ◽  
Three Dimensional ◽  
3D Culture ◽  
Stem Cell Differentiation ◽  
Culture Conditions ◽  
C2c12 Cells ◽  
Dependent Manner ◽  
Physical Force ◽  
Myod Gene ◽  
2D And 3D

Abstract Among many factors of controlling stem cell differentiation, the key transcription factor upregulation via physical force is a good strategy on the lineage-specific differentiation of stem cells. The study aimed to compare growth and myogenic potentials between the parental cells (PCs) and the 1-day-old C2C12 spheroid-derived cells (SDCs) in two-dimensional (2D) and three-dimensional (3D) culture conditions through examination of the cell proliferation and the expression of myogenic genes. The data showed that 1-day-old spheroids had more intense expression of MyoD gene with respect to the PCs. The proliferation of the SDCs significantly higher than the PCs in a time dependent manner. The SDCs had also significantly higher myogenic potential than the PCs in 2D and 3D culture conditions. The results suggest that MyoD gene upregulation through cell-cell contacts is the good approach for preparation of seed cells in muscle tissue engineering.

Three-dimensional tissue model in direct contact with an on-chip vascular bed enabled by removable membranes

Lab on a Chip ◽  
2022 ◽  
Author(s):  
Yoshikazu Kameda ◽  
Surachada Chuaychob ◽  
Miwa Tanaka ◽  
Yang Liu ◽  
Ryu Okada ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Direct Contact ◽  
Three Dimensional ◽  
Culture Period ◽  
Tissue Model ◽  
Vascular Bed ◽  
On Chip

Three-dimensional (3D) tissue culture is a powerful tool for understanding physiological events. However, 3D tissues still have limitations in their size, culture period, and maturity, which are caused by the...

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