3D Cell Transfection Kit 24 wells

In stock
SKU
3D-DNA24
150,00 €
3D Cell Transfection Kit 24 wells
£140 / €150

Supplier
BioCellChallenge
Cat. no.
3D-DNA24
Kit size
1 kit
Links
BioCellChallenge
Protocol
data
Brochure

Features

  • high delivery efficiencies of plasmid DNA into 3D cultured cells
  • includes a sterile plate containing 3D Cell Culture Scaffolds
  • In vivo-like cell culture environment
  • One step « Transfect & Seed » protocol
  • Compatible with inverted light microscope

The 3D Cell Transfection Kit is based on a unique in vitro transfection technology which allows researchers to achieve high delivery efficiencies of plasmid DNA into 3D cultured cells. This innovative product includes a sterile plate containing 3D Cell Culture Scaffolds (3D Biotek, LLC) and an especially designed 3D Transfection Reagent (BioCellChallenge, SAS). With this kit, researchers can now perform extended 3D transgene expression studies in cells grown in physiological-like tissue environments.

3D Cell Culture Scaffolds are made from polystyrene, the same material as traditional tissue culture plates. The combination of polystyrene’s transparency and the porous structural design of 3D Cell Culture Scaffolds allows researchers to monitor cell growth and transfection efficiency under an inverted light microscope.

3D Cell Culture Scaffolds within the 3D Cell Transfection Kit have increased surface areas compared with 2D cell culture plates. As a result, more cells can be transfected on 3D Cell Culture Scaffolds while using the same size cell culture plates.

3D Cell Culture Scaffolds will not absorb cytokines, growth factors, or any other molecules. Therefore, molecules secreted by the transfected cells can easily be separated or recovered from culture medium without extensive separation steps.

Protocol: A 3D Cell transfection protocol is provided for a 96-well scaffold. A table is provided to adapt it to the other scaffold size

    1. Dilute 0.5 µg of DNA in 17 µL of serum free medium.
    2. Add 3 µL of 3D Transfection Reagent to the diluted DNA solution and mix the solution by pipetting.
    3. Incubate 15 minutes at room temperature.
    4. During this incubation time, spin down 200,000 healthy cells for 5 min at 1,000 rpm in a microtube and discard supernatant.
    5. Add the 20 µL of 3D Transfection Reagent / DNA mixture onto the pellet of cells, mix gently by pipetting and slowly pipette this 20 µL suspension cell onto the top center surface of the 3D cell culture scaffold.
    6. Incubate plate at 37°C in a CO2 incubator.
    7. After 3 hours of incubation, slowly add 180 µL of complete medium (containing serum) and return the plate to the CO2 incubator. Ensure that the scaffold is sitting at the bottom of the well. If not, gently push it down with a pipette tip.
    8. After 24 hours, carefully aspirate the medium around the scaffold and then flip and transfer it into a new well with sterile forceps. Slowly add fresh medium into the well and make sure that the scaffold is sitting at the bottom of the well.
    9. Monitor the 3D cell transfection under an inverted light microscope. Analyse the transgene expression or continue the transgene expression for later analyses, according to your experimental protocol.
  • Kit Components
3D Transfection Reagent 200 µl
24-Wells scaffold 1


  • Storage and Handling
The plate containing the 3D Cell Culture Scaffolds can be stored at room temperature. 3D Transfection Reagent is stable for at least 1 year at 4°C.
In vivo-like cell culture environment.

Figure 1: Cells growing on 3D InsertsTM (3D Biotek, LLC) were transfected with a plasmid DNA encoding GFP and the 3D Transfection Reagent (BioCellChallenge, SAS). Protein expression was monitored by fluorescence microscopy 24h to 72h post transfection.

Fibroblast cells transfection.



NIH3T3 cells transfection


  • It has been well documented that cell culture in tissue culture plates and flasks does not mimic the in vivo cell growth. Therefore, the cell expansion and cell-drug interactions under 2D are not appropriate in vitro models. 3D cell culture technique, on the other hand, offers a better cell culture environment because it is one step closer to the in vivo cell growth environment.
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