Feature of NanoCulture Products
NanoCulture Plate / Dish is serialized from 24, 96, 384-well plate to 35 mmφ dish. Variety of well sizes is applicable for different experiments, from HTS to large scale preparation of spheroids.
NCP (24, 96, 384 well) and NCD (35 mmφ）
Advantage of NanoCulture series
|Ready-to-Use||Same as conventional monolayer culture plates. After opening the package, just add cell suspension into wells.|
|Good Microscopic Observation||Capable of parfocal observation for all spheroids within the view field. Can be measured directly with fluorescence/luminescence plate readers.|
|Uniform Production Lot||Coating reagents free. No biological materials. Made of plastic material, with almost no lot-to-lot variation.|
|High Cell Viability||Cells obtain high proliferation capacity by adhesion signal. Also, cells form spheroids while migrating, which cause highly viable cells attached to the plate form spheroids.|
|Rapid Spheroid Formation||Capable of microscopic observation of spheroids by day 3 after seeding.|
|Large Scale Preparation of Spheroids||35mmφ dish with same system as plates is available. Capable of large scale preparation of spheroids with enlarged culture area.|
Spheroid morphology depend on the cells character
NCP has black casing and clear flat bottom, made with a high transparent plastic film, which is suitable for imaging and compatible with automated lab equipment. Further, the spheroids are attached to the bottom film, which can be focused for obtaining clear images under both bright-filed and fluorescence. Cells form spheroid according to their character. It is considered that E-cadherin is key molecule for spheroid formation. Epithelial cancer cell expressing E-cadherin in high level and they form compact spheroid indicating tight cell-cell attachment. By contrast, which cells form loose aggregation like a grape shape, i.e. cell cluster, expressing low level E-cadherin.
Spheroids with smooth surface
Spheroids with rough and bumpy surface
Grapevine-like clusters spheroids
Observed with a confocal fluorescence microscopy after CalceinAM stain
Observed with a Transmission Electron Microscope
- Human breast adenocarcinoma cell line MCF7 forms a hollow/lumen on NCP
Data provided by: Prof. Hideo Namiki, Waseda University
Comparison of NCP with other 3D cell culture methods
NCP/NCD are the scaffold type 3D cell culture system. The products have advantages in point of proliferation and viability of cell, and overcome flaws of scaffold type 3D culture system in high-throughput screening, e.g. lot-to-lot variation, cumbersome procedure, difficulty of imaging.
Cells can actively proliferate in NCP as well as in Matrigel (scaffold type) while very low proliferation activity in low adhesion plates (scaffold-free type)
Proliferation curve of HepG2
ATP concentration of cultured HepG2 were measured at day 1、5、7、10 and 14. HepG2 cultured in NCP have high proliferative activity as well as in Matrigel. In contrast, proliferation activity of the cells in low adhesion plates is low level.
Spheroids cultured in NCP have high cell viability compared with low adhesion plate. One of the reason is that low penetration of nutrient and/or oxygen inside the spheroid. It may be considered that cells keep their viability in NCP because the spheroid size doesn’t become excessively big.
Images of HT29 cells stained with CalceinAM
HT29 cell line was seeded on NCP (96 well plate) at 1×104 cells/well. After 7 days culture, spheroids were dispersed into single cells, and then stained with CalceinAM. Almost all cells have green signal that represents high cell viability on NCP.
Low attachment plate (round bottom)
Low attachment plate (flat bottom)
Images of HepG2 spheroid stained with EthD-1
Dead cells are indicated in red. In NCP, living cells attach to the bottom of plate and form spheroids. Dead cells are not included in spheroids and floated in culture sup.
Comparative Table of NCP/NCD and Other 3D Cell Culture Methods
|Uniform Manufacturing Lot||＋＋＋||–||＋＋＋|
|High Cell Viability after 1-Week Culture||＋＋＋||＋＋＋||＋|
|Fast Cell Growth||＋＋＋||＋＋＋||＋|
|Fast Spheroid Formation||＋＋＋||＋||＋＋＋|
|Wide Variation for Well Sizes(from mass culture to HTS)||＋＋＋||＋＋||–|