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  3. FAQ NCP – Introduction




Q.101 – What is the use of NanoCulture Plate?
NanoCulture Plate (NCP) is a cell culture plate used for three dimensional cell culture. Cells grown in NCP can form multi-cellular clusters (spheroid) after seeding cells like conventional monolayer cell culture plates. No specialist equipment is required. Microscopic observation can be made with clear well-bottom film, while culturing. Also, black upper structure allow fluorescence imaging.
It is used for various applications, such as 1) evaluation of intracellular changes during 3D cell culture, 2) anticancer drug screening (compatible for HTS), 3) evaluation of knockdown effect with siRNA, 4) hepatotoxicity screening, 5) differentiation of stem cells, etc.
Q.102 – What is three-dimensional (3D) cell culture? What is the advantage?
Three-dimensional (3D) cell culture is a method to form three-dimensional multi-cellular cell cluster (spheroid). Cell morphology is different compared to conventional culture method using polystyrene culture ware (two-dimensional (2D) culture, monolayer culture). Multi-cellular spheroid is known to have closer characteristics to in vivo compared to monolayer cultured cells.
Q.103 – How do cells on NCP form spheroids?
Cells seeded on NCP attach to the ultrafine processed mesh structure on the culture surface, grown in a spherical morphology. Cells migrate on the microstructure, and continuously contact/adhere to each other and form multi-cellular clusters (spheroids). Spheroids grow larger with proliferation at the same time.
Reference: Technology
Q.104 – What is the difference between other three-dimensional cell culture methods?
Two of the most popular 3D cultures are, by forming spheroids 1) on matrix or gel, or 2) suspended with Ultra-low attachment microplates. NCP is a ready-to-use microplate, which can form spheroids with a standard culture medium, and can be easily observed by standard microscope through the transparent bottom film, and has following advantages.
  • ・No foreign unknown materials
  • ・Uniformly grown spheroids
  • ・Compatible with High-throughput
  • ・Easy to observe through transparent bottom film
  • ・High cell viability (compared to Ultra-low attachment microplates)
  • ・Easy to collect cells
  • ・Easy to change medium (spheroids attach to the plate, compatible with fluid medium)
Q.105 – What material is NCP made of?
NCP is made of synthetic polymer, with low attachment surface. By imprinting micro-structure patterning, cell adhesion is controlled to provide suitable environment for cells to form spheroids.
Q.106 – Is the culture surface coated?
No, it is not coated with any chemical substances, protein, gel, etc.
Q.107 – What types of NCP are available?
SBS format micro-titer plates (384-, 96- and 24-well) and 35 mm dish are available. Also, two types of adhesion (high/low) and different imprint pattern (honeycomb/square) can be chosen.
*High-binding grades are produced on order.
Q.108 – Are the spheroids attached to the culture surface (bottom film)?
Spheroids are attached to the bottom film, however, its adhesion is not strong. Cells contacting the culture surface and grab the micro-structure as a scaffold.
Q.109 – How can the spheroid formation be confirmed?
It can be confirmed by microscopic observation.
Q.110 – Does 3D culture by NCP require special kind of equipment?
No, it does not require any special equipment. It can be used with same experimental equipment or device as the standard monolayer culture. We recommend to pre-incubate the plates before culturing, and it is useful if you have a microplate centrifuge. Also, it is useful to apply microplate compatible colony counter, which can measure spheroid size and its distribution.
Q.111 – Can NCP be reused?
No, you cannot reuse the used NCP wells. The unused wells within the plate can be reused for another experiment, by making aseptic handling (such as by covering the wells with plastic seal).
Q.112 – Can NCP be sterilized by autoclaving?
No, please do not autoclave NCP as it may cause deformation. NCP is sold in sterilized package. To maintain sterility, please make sure not to open the lid of NCP outside the clean bench.
Q.113 – Is NCM a must-use medium for 3D culture using NCP?
No, it is not a must for NCP spheroid culture. Spheroids can be formed on NCP by using standard culture medium with 10% FBS content.
Q.114 – What kind of medium is NCM?
Two types of NCM are available, 1) M Type, and 2) R Type with enhanced spheroid formation. Both medium are sold in a set of basal medium added with antibiotic and FBS. FBS is a lot-checked by spheroid formation on NCP. Normally, 10% FBS content is recommended, however, can be changed to a suitable concentration by monitoring the spheroid morphology.
Q.115 – What is the difference between Spheroid Dispersion Solution and Spheroid Lysis Buffer? And, when should it be used?
Spheroid Dispersion Solution is a solution which will disperse spheroids into single cells enzymatically (not trypsin). Single cells obtained by the dispersion of spheroids can be used for successive subculture, or Flow Cytometry analysis, etc.
Meantime, Spheroid Lysis Buffer lyse spheroids. This solution completely destroy the cells, which cannot be used for successive subculture. Further, as it is a very strong detergent, it cannot be used for experiments where protein conformation reservation required. It can be used for PicoGreen Assay (method to measure viability by DNA content).
Q.116 – What cell types can form spheroids on NCP?
It is for adherent cells. SCIVAX confirmed spheroid formation using various cell types, mostly cancer cell lines. Please contact us for updated information as we are continuing to evaluate various cell types.
Floating cells, such as blood corpuscle cells may be difficult to form spheroids, however, if it has an adhesive property, it may be worth trying.
Reference: Spheroid List
Q.117 – Is it possible to request spheroid culture on contract basis?
Yes, it is possible. SCIVAX is providing contract spheroid culture service. We also offer on-sight training. Reference: Contract Service