Human amniotic fluid stem cells (University College, London)

Amniotic fluid samples were collected by amniocentesis (gestational age 14–25 wk) during routine prenatal testing. Samples were then centrifuged at 1500 rpm and cell pellets were resuspended and seeded in Chang medium comprised of α-MEM supplemented with FBS (15% ), glutamine (1%),and penicillin/streptomycin (1%), Chang B (18% ) and  Chang C (2%). After three days of incubation, nonadherent cells and debris were discarded and the adherent cells were cultivated  to near confluence. Adherent cells were immunomagnetically sorted for the stem marker c-kit, using a mouse monoclonal anti-c-kit (CD117) antibody and the anti-mouse IgG Cellection Dynabeads M-450 antibody. c-kit-positive cells were reseeded and expanded in Chang medium.