Questions and Answers

Recent answers:

Q: Are there any published papers concerning the use of alvetex®?
A:

There are numerous peer-reviewed scientific articles available describing the development and application of alvetex® (see section on publications). Scientists at reinnervate continue research into novel applications for this technology and publish and present their data when appropriate.

Q: Can I coat alvetex® with known proteins and reagents?
A:

Yes. Most standard coating methods for cell culture plastic are compatible with alvetex® including the use of extra-cellular matrix proteins and poly D / L lysine etc, for more information, please visit the technical support section of the web site

Q: Can I remove my cells from the scaffold?
A:

Yes. Partial cell recovery is possible. Cells can be removed from the scaffold using a combination of trypsin and mild agitation. For more information, please visit the technical support section of the web site

Q: Can I see cells using standard light microscopy?
A:

Alvetex® has been developed to enable 3D cell growth and users can generate micro-slabs of tissue. This means that standard light microscopy is not suitable for the visualisation of 3D cells growing in alvetex®. More sophisticated methods such as confocal microscopy can be used to see cells cultured on alvetex® (see image library and technical brochure). Depending on the cell density, it is possible to pick out individual cells growing within large voids. Alvetex® is very well suited to tissue fixation, processing and imaging techniques such as IHC, histological staining, immunofluorescence, SEM and TEM

Q: Can I use more than one scaffold within the same well?
A:

There are several options available to scientists who wish to experiment with co-culture and cell invasion assays.

  • Two or more cell types can be seeded into one alvetex disc simultaneously; One cell type can be grown in alvetex and then, at a later time, a second cell type can be seeded onto the top of the alvetex® disc;
  • Cells can be grown in 3D at the bottom of an alvetex® 12 well plate and different cells, growing in an alvetex 12 well insert, can be grown above the first set of cells. The 2 cells types may therefore be able to exert a paracrine effect upon each other;
  • It is possible to seed and grow two different cell types inside 22mm alvetex discs. One of the alvetex® discs can then be moved and placed on top of the other to bring the 2 sets of 3D cells into contact with each other.
Q: Does alvetex® come ready to use?
A:

Like standard cell culture plastic-ware, alvetex® is ready to use from the moment it is unwrapped. For more information, please visit the technical support section of the web site.

Q: Does alvetex® degrade?
A:

Alvetex® is inert and does not degrade during standard cell culture practice.

Q: Does alvetex enable 3D Growth?
A:

Cells grown in alvetex® adopt a 3D geometry. Cells grow in contact with adjacent cells forming a thin 3D layer. See 'image library' and 'publications' for scientific evidence.

Q: Does the material auto fluoresce?
A:

Alvetex® does not auto fluoresce under the standard conditions we have tested for fluorescent microscopy.

Q: Has alvetex® been treated in any way?
A:

Alvetex® has been sterilised using standard techniques.

Q: How do I seed cells onto alvetex®?
A:

Reinnervate provides guidelines for the seeding of some specific cell types onto alvetex®. For more information, please visit the technical support section of the website (insert link). Whilst we are developing more cell specific protocols all the time, it may be necessary to carry out a small amount of optimisation of your cells. If you need help or advice, please call or email us and we will be pleased to advise you.

Q: How is alvetex® supplied?
A:

Alvetex® is supplied packaged either in multi-well culture plates or in individual blister packed well inserts designed to fit all standard 6 and12 plates. New formats of alvetex® will be available soon (including 96 well plates). If you want to be the first to know about future product releases, please sign up for our e-mail alert service

Q: How much culture media do I use?
A:

For any given culture vessel type, cultures grown in 3D contain more cells than their 2D counterparts. Accordingly, the metabolic demands on that media are generally higher in 3D. In a standard plate, the vessel should be filled with media to its maximum recommended level. To address this issue, reinnervate has developed a series of new devices to enable users to culture cells in 3D using alvetex® with larger volumes of culture media. For more information, please visit the technical support section of the web site

Q: How often do I need to change the culture medium?
A: This depends on the nature of the cells grown in 3D culture. Within a standard culture vessel, a 3D culture will require more frequent media changes compared to its 2D counterpart. However, this requires optimisation for the cell type concerned. reinnervate has developed a series of new devices to enable users to culture cells in 3D using alvetex® with larger volumes of culture media. This equipment allows fewer media changes and maintains culture media quality. . For more information, please visit the technical support section of the web site
Q: Is alvetex® available in a well insert?
A:

Yes - alvetex® is currently available in well inserts as 6 and 12-well formats.

Q: Is alvetex® compatible with other technologies?
A:

Alvetex® is a versatile material that can be adapted for use in a broad variety of applications. We encourage investigators to explore its use in alternative and novel areas. Please enquire should you require advice about your particular application.

Q: Is alvetex® easy to use?
A:

Alvetex® is very straightforward to use. It has been developed specifically for routine 3D cell culture and can be used alongside existing cell culture products. Full instructions are provided.

Q: Is alvetex® re-useable?
A:

No, like any plastic consumable product for cell culture, it is not recommended to re-use alvetex®.

Q: Is alvetex® sterile?
A:

alvetex® is supplied as a sterile packaged product ready to use.

Q: What cell types have been tested?
A:

A broad range of cell types have been grown on alvetex® scaffolds. These include hepatocytes, keratinocytes, fibroblasts, stem cells, osteoblasts, various epithelial cells, neurons, astrocytes, primary cells and tissues, and various other cancer cell lines.

Q: Can I use alvetex® for co-culture experiments?
A:

There are several options available to scientists who wish to experiment with co-culture and cell invasion assays.

Two or more cell types can be seeded into one alvetex® disk simultaneously;

Once cell type can be grown in alvetex® and then, at a later time, a second cell type can be seeded onto the top of the alvetex® disc;

Cells can be grown in 3D at the bottom of an alvetex® 12 well plate and different cells, growing in an alvetex® 12 well insert, can be grown above the first set of cells. The 2 cells types may therefore be able to exert a paracrine effect upon each other;

It is possible to seed and grow 2 different cell types inside 22mm alvetex® discs. One of the alvetex® discs can then be moved and placed on top of the other to bring the 2 sets of 3D cells into contact with each other.

Q: Can I use alvetex® for cell invasion assays?
A:

Whilst this is not something we have yet tried, it should be possible to set up this type of test. Cells, matrix proteins or other compounds can be seeded into alvetex and then the cell of interest seeded onto the top of alvetex®. Standard tissue processing, fixing and staining methods could then be employed to monitor cell invasion.

Q: What evidence is there that alvetex® works?
A:

Scientists at reinnervate have thoroughly exemplified the use and application of alvetex®. The image library provides evidence of 3D cell culture throughout the scaffold. Much of the data generated using alvetex® has been published in peer reviewed scientific journals and/or presented at various inter/national scientific meetings (see publications section).

Q: What general techniques can I use to analyse my samples?
A:

Alvetex® is compatible with a broad range of general cell and molecular techniques. These include various biochemical assays, histology (tissue processing, fixation, embedding and sectioning), fluorescence microscopy, immunocytochemistry, in situ hybridisation, electron microscopy, bright field and phase microscopy; cryostat sectioning, isolation of viable cells for passaging; extraction of nucleic acid and total protein, etc.

Q: What is alvetex® made of?
A:

Alvetex® is made of cross-linked polystyrene.

Q: What is the porosity of the material?
A:

The voids have an average diameter of 40 µm and inter-connects have an average diameter of 13 µm. Alvetex® is extremely porous to enable cells to penetrate and grow throughout the material.