I normally starve cells for 24 hours in serum free media and then add my drug in serum media, scratch and add this treatment media and follow the healing. In case of HaCat I starved the cell in 1% FBS containing media for 24 hours, however the healing was not observed even in control. Please share your experience with this cell line for scratch assay.
I did a search on SciGine and all the results seem to indicate that HACAT need to be in 10% FBS prior to a wound healing assay. Take a look at the attached method and feel free to do some searches as well.
http://scigine.com/method.php?ID=231429
We published some research on that. I hope it helps.
Amniotic Membrane Modifies the Genetic Program Induced by TGFß, Stimulating Keratinocyte Proliferation and Migration in Chronic Wounds
Antonia Alcaraz,
Anna Mrowiec,
Carmen Luisa Insausti,
Ángel Bernabé-García,
Eva María García-Vizcaíno,
María Concepción López-Martínez,
Asunción Monfort,
Ander Izeta,
José María Moraleda,
Gregorio Castellanos,
Francisco José Nicolás
PLOS
Published: August 18, 2015
http://dx.doi.org/10.1371/journal.pone.0135324
In my experience, starving the HaCaT in serum free media does not do well with the scratch assay. I have done a side to side comparison with HaCat's that were starved and the ones that were maintained in 10% FBS. The cells in 10% FBS migrated and closed the scratch area as expected in 48 hours.
However, I must say, the rate of migration for the HaCaT is slower than some other cell lines. I am yet to determine why.
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