FREQUENTLY ASKED QUESTIONS
Q: Can I seed cells in this plate as I would in a normal 6-well plate? Should I change the cell density?
A: Yes, we recommend that our customers seed cells using the same density that they do on conventional 6-well plates.
Q: What is the depth from the nanopatterned surface to the plate bottom?
A: This dimension is about 0.7 mm.
Q: Is the plate bottom parallel to the nanopatterned surface or is there a cone-shaped slope in order to collect cells to the nanosurface?
A: The plate bottom is parallel to the nanosurface. However, there will be no problem seeding the cells on the surface if the protocols we recommend are followed.
Q: Can I use the same volume (of medium) as usual?
A: Yes. The working volume is 3 mL. Also, amount and duration for cell media change should be the same as with normal 6-well or single-well culture.
Q: Do cells adhere not only to the nanopatterned surface but also to the plastic plate bottom?
A: Ideally, no if you follow the protocol provided. However, there is a possibility that cells can attach on to plastic plate bottom if you don’t follow the protocol.
Q: Is there any type of pre-wash that must be performed prior to seeding the dishes?
A: No, ANFS is already gamma sterilized and plasma treated to promoted cellular adhesion. However, researchers may choose to deposit additional surface treatments depending on their exact cell type. Fibronectin, collagen, and PLL are all common surface treatment choices.
Q: Why 800 nanometers?
A: We arrived at the 800 nanometer 1:1 dimension after considerable research and development work. We scanned through many different dimensions, ranging from tens of nanometers to a few microns, and found that 800 nm was optimal for promoting cellular maturation and even differentiation of many cell types. This is likely because topographical features of this dimension resemble native collagen fibers in the extra cellular matrix. Additionally, we define 800 nm as a standard to give researchers a controlled parameter; unlike electrospun fibers, which can vary significantly in dimension, our nanopatterned surfaces have consistent, defined dimensions, so your studies are controlled.
Q: What protocols do you have for the use of the dishes. In the literature, several different seeding densities were used. What do you recommend?
A: As with any cultureware, optimal seeding density on ANFS can vary according to cell type and the purpose of the experiment. However, we recommend researchers use exactly the same cell density that they use for normal 35mm dishes, 6-well dishes and so on. Protocol information is provided on the ANFS product pages.
Q: What is the shelf life of the dishes?
A: Shelf life is generally indefinite, but use is recommended within 2 years.
Q: What is your suggestions for culturing undifferentiated primary cells?
A: This depends on cell source and the stage the cells are in. However, we recommend the same protocols for stem cell-derived somatic cells
Q: What is your suggestion for culturing skeletal muscle cells?
A: We recommend similar protocol for SMCs as we do for cardiomyocytes. For coating, however, we normally used matrigel, gelatin or Collagen I.