FAQs

Precautions and Disclaimer
This product is for R&D use only, not for drug, household, or other uses.

Storage/Stability
dPGA is highly stable and can be stored long-term at 4 °C.

Stock Preparation
Under sterile conditions, dilute dPGA to a 1 mg/ml stock solution with sterile ddH20. A sterile 1
mg/ml stock solution can be stored at 4 °C long term (up to 2 years).

Dendrimer Coating Protocol
Cell culture:
When using dPGA as a coating to promote cell-substrate attachment, the optimal conditions
should be determined for each cell line and application.
1. Under sterile conditions, dilute the 1 mg/ml dPGA stock solution to a final concentration
of 50 μg/ml (1:20 dilution) in sterile 1XPBS. The final concentration of the coating solution
may vary from 10-100 μg/ml. We recommend using a concentration of dPGA that is the
same as the concentration of poly-lysine in similar protocols.
2. Add the dPGA coating solution to your culture vessel and incubate for at least 1 hr at 37°C
(we recommend placing them in the CO2 incubator for incubation).
3. Following coating, remove the solution by aspiration and wash 3x with sterile 1XPBS.
4. After the last wash, seed the cells or coat with a second layer of Laminin or Matrigel (see
Step 5 below). Note: Leave the dPGA coated plates in 1XPBS until use.

Second layer coating with Laminin or Matrigel
5. After step 3 (above), the dPGA coated substrate may be coated with laminin or Matrigel.
For example: 20 μg/mL laminin solution or Matrigel in DMEM/F-12. Incubate the plates
for 1-2 h at 37 °C or overnight at 4 °C.

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dPGAs (dendritic polyglycerol amine) are a family of polymers with a highly branched dendritic structure composed of glycerol monomers with a portion of the glycerol hydroxyl end groups substituted with amine functional groups. dPGA presents a high density of positive charges at neutral pH, but unlike poly-lysine and poly-ornithine, does not contain peptide bonds and is therefore highly resistant to proteolysis.

dPGA is a synthetic polymer. It is not a protein and therefore, in contrast to PLL,PDL, PLO, fibronectin and other protein base coating substrates, will not be degraded by protease overtime ensuring stability of the cell culture coated surface.

There’s a lot of issues raised from researchers around the globe about how difficult primary neuronal cultures can be. Some ask why are my primary neuronal cultures dying in culture? How can I keep them healthy, long term? Others observe clumps in neurosphere/aggregate-like days after seeding, what's wrong? 

While many factors should be taken into consideration when facing cell culture challenges, the surface treatment of your vessels is an important one.

Our data shows that dPGA coated surfaces can support long-term cultures of primary cortical neurons up to 3 months with a higher cell density, preservation of neuronal physiology characteristics and prevention of cell clumping/aggregates compared to PLL, PDL and PLO.

Cells in culture secrete proteases that degrade polypeptide based substrates like poly-lysine and poly-ornithine. Over time, this results in a healthy culture degrading its own substrate. On a sub-optimal or degraded culture substrate, cells form clumps because they adhere better to each other than to the poor substrate. The dPGA polymer is not a protein and not degraded by proteases, resulting in massively enhanced stability as a cell culture substrate.

Coating your vessels with dPGA for primary neuronal cultures is easy and comparable to standard PDL coating protocol.

Please refer to our protocol in the FAQ section or available as a PDF format on this page https://dendrotek.ca/products/dpga

dPGA can be stored at 4C long term. You can use it on glass or plasticware surfaces and culture plates can be coated in advance, dried and stored at 4C up to 3-4 days for later seeding of your cells.

No. Using dPGA involves very simply replacing poly-lysine and poly-ornithine in your existing protocol for coating the cell culture substrate.

Yes, You can pre-coat you plates with dPGA.

After the last PBS rinse, remove the rinse and add sterile 1X PBS. Wrap your cultureware in parafilm and store at 4C, wet, for up to 2 weeks. Make sure to preserve sterility of your vessels.

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