NanoCulture Plate

MBL International, together with ORGANOGENIX (formerly SCIVAX Life Sciences), now offers the NanoCulture Plate for 3D cell culturing. The NanoCulture Plate/Dish (NCP/NCD) is engineered with an evenly patterned nano-scale structure on a propietary plastic film attached to the bottom of the plate/dish. The pattern mimics the extracellular matrix and allows cells to migrate across the plate and form healthy spheroids.

In addition, the pattern eliminates the need for any kind of biological/gel or matrix, allows for easy handling, and is fully compatible with lab apparatus, imaging apparatus, and high throughput screening systems. The plate is also compatible for drug screening, co-culture assay, EMT assay, hepatotoxicity assay, ADME assay, primary cell culture, stem cell culture and differentiation.

It is well known that scaffold is very important for 3D cell culture to mimic tissue micro environment. However, working with gels are always a cumbersome procedure in terms of HTS or any kind of assay development. Therefore, researchers can use our plate instead of a gel platform and receive better biological relevance data compared to other plates, because the cells are free to move on NCP and forming spheroids. As a result, no gravity force or external materials are needed.

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NanoCulture Technologies for 3D Cell Culture                                                                                                                

Totally gel-free but scaffold-type 3D cell culture method – Micro-structure acts like a scaffold for cells

NanoCulture Plate/Dish (NCP/NCD) is engineered with an evenly patterned nano-scale structure on a proprietary plastic film which is the plate bottom surface. This pattern mimics the extracellular matrix and supports the cell to form cell spheroids.  This nano-scale structure controls the adhesion of cells to the film, and enhances cell migration and proliferation while cells form spheroids.

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Nano-scale patterned well-bottom film                                                                                                        

Structural color is observed, due to the interference of light by nano-scale pattern.

 

 

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HeLa cell spheroid attaching to the nano-scale structure of film                        

Several cells constituting a spheroid, contact the well-bottom film,  grip and  attach to nano-scale pattern using pseudopodium.  Data provided  by: Prof.  Hideo Namiki, Waseda UniversitySpheroid formation mechanism on NanoCulture Plate/Dish (NCP/NCD).

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Cells seeded on a NCP/NCD, grip and attach to the nano-scale pattern on a culture surface.  Cells start migrating randomly and dynamically as they do not adhere strongly to the base film thanks to the micro-structure, unlike conventional monolayer cell culture.  Cells contact and adhere to each other while migrating on the surface,  and  form spheroids, accompanied by proliferation at the same time.  Spheroid growth reaches a plateau as the culture period  progresses.

 

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Chronological change of spheroid size of HepG2 cells

Box plot of spheroid diameter obtained from HepG2 cells cultured for 7 days on NCP (96 well plate) at 10,000 cells/well. Spheroids keep growing as culture period progresses, while it reaches to a plateau in approximately one week.

Format & Advantages of NanoCulture Series

Assay-friendly well-sizes, optimized micro-structure pattern, and cell adhesive.

NCP and NCD are available as a 24, 96, 384-well plate or a 35 mmΦ dish.  A variety of well formats is applicable for different experiments, from HTS to large scale preparation of spheroids.

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NCP (24, 96, 384 well) and NCD (35mmΦ)

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Structure of culture surface – Left: Microsquare (MS), Right: Microhoneycomb (MH)

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There are two patterned micro-structures (Microsquare/Microhoneycomb) and two types of cell adhesive in bottom film. We have evaluated more than 80 cell lines, and confirmed of pattern “preference” with approximately 10% of the cell types.  (These results are exhibited in our website. For more information, please contact us, or test both patterns of NCP/NCD with your cell types).

Advantages of NanoCulture Plate/Dish

Easy 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. Fully compatible with HTS imagers, confocal and convention

Reproducibility > Coating reagents free. No biological materials. Made of plastic material, with almost no lot-to-lot variation.

High Cell Viability > Nano-scale scaffold allows natural migration and formation of spheroids resulting in high proliferation compared to  scaffold free 3D platforms.

Natural Spheroid Formation > Scaffold system eliminates the need for any external forces in spheroid formation.

Large Scale Preparation of Spheroids > 35mmΦ dish with the same technology is available.  It is suitable for large scale 3D cell culture.

Comparative Table of NCP/NCD and Other 3D Cell Culture Methods

NCP/NCD Scaffold Scaffold-Free
Easy Handling +++  ++
Excellent Observation(time-lapse imaging) (fluorescent imaging) +++ +  +
Uniform Manufacturing Lot +++  +++
High Cell Viablility after 1-week culture +++ +++  +
Fast Cell Growth +++ +++  +
Fast Spheroid Formation +++ +  +++
Wide Variation for Well Sizes(from mass culture to HTS) +++ ++  –

Spheroid morphology on NanoCulture Plate/Dish

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Spheroid morphology is totally depending on cell characteristics (migration, proliferation, intracellular adhesion molecules expression, or degree of differentiation). Typically, it can be classified into 4 types.

Spheroid culture medium (M-type/R-type) and Medium Supplement

Though cells can form spheroids in NCP with standard growth medium, we provide two types of culture medium.  NanoCulture Medium (NCM) is optimised for spheroid formation, in combination with lot-checked serum.  In case of insufficient spheroid growth with M-type medium, it may improve by using R-type medium.  We can also supply medium supplement, NanoCulture Medium Supplement (10x).  FBS is not included in this product, as you can prepare a spheroid culture medium with your own FBS.

Spheroid Lysis Buffer and Spheroid Dispersing Solution

Spheroid Lysis Buffer (SLB) dissolves spheroids into Lysate.  It can be used for DNA quantification for viablility measuring.  Spheroid Dispersion Solution (SD4x) enzymatically disperses spheroids to single cell.  The cultured single cells can be used for cell counting, flow cytometry and subculture, etc.

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Tumour Organoid Culture Kit

The need to harvest primary cancer cells from tumour in a 3D system arises from our desire to understand the patient specific tumour biology.  It is widely accepted that data obtained from primary cells is not only preferred, but most relevant when trying to study physiological interactions.  By culturing primary cancer cells on NCP, stromal cells do not cause overgrowth, but form co-cultured spheroids with cancer cells.  This kit is enabled to test and evaluate anti-cancer drug sensitivity from a small amount of specimen (>300 mg). The kit consists of three major components.  1) Primary Culture Medium, containing growth factor supplements, 2) Dissociation Solution and tools, and 3) NanoCulture Plate.

Caltag Medsystems are the UK distributor for MBL International products. Please contact our technical support team to discuss your requirements further.

 

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