PA-1 differentiation takes between 7 and 14 days. In 7 days cells already present differentiated characteristics, however 14 days of differentiation ensures the highest level of differentiation. PA-1 cells differentiate into an ectodermal linage (fibroblast, neurons and glia).
PA-1 are able to differentiate under different culture media conditions, however, the most optimal condition is the differentiation with 10 % KOSR (KO Replacement Serum) and 1 µM RA (Retinoic Acid).
As PA-1 cells become differentiated, they are more delicate and gets harder to passage them succesfully. Due to this, when PA-1 differentiating cells need to be passed, it is neccessary to create a favorable surface for them to attach. For this reason culture dishes must be treated with matrigel for at least 1 hour prior cell seeding.
During differentiation, culture media with 10% KOSR and 1 µM RA should be changed every other day.
PA-1 Differentiation culture media
- 450 ml 450 ml MEM + GLUTAMAX (GIBCO X10, 42360032 – 42360099x10)
- 50 ml KnockOut Serum replacement medium (10%, KOSR, Invitrogen, 10828028)
- 1µM of retinoic acid (RA, trans-only R2625-100MG, Sigma)
- 5 ml of 1% Penicilin/Streptomicin 100 ml (100x). (GIBCO, 15140122)
- 5 ml of 1% MEM-NEAA (GIBCO, 11140-035)
- Only if MEM doesn't include Glutamine: 1% GLUTAMAX-1 SUPPLEMENT (GIBCO, 35050-038) o L-GLUTAMINA 200mM (100x) 20ML (GIBCO, 25030-032) (in case the media doesn’t have L-Glutamine)