Mouse embryonic stem cells (ESCs) from strain Sv/129 were cultured according to a protocol described in our previous publication [2, 3] . The mouse ESCs came from Xuanwu Hospital (Beijing, China) and were maintained with mitotically-inactivated mouse embryonic broblasts (MEF). When passaged, ESCs were seeded onto culture dishes coated with 0.2%
gelatin (Sigma-Aldrich, St. Louis, MO, USA; Cat# G1393), and LIF was added to maintain the ESC characteristics. The ESCs were cultured in
Dulbecco's modi ed Eagle medium without folic acid (Sigma-Aldrich; Cat# D2429). The cells were divided into three treatment groups according to folate concentration: folate-free group (ESC/FF, 0 mg/l), folate-de cient group (ESC/FD, 0.5 mg/l) and normal group (ESC/FN, 4 mg/l). The cells were cultured in a humidi ed environment containing 5% CO 2 , at 37°C. When approximately 70-80% con uency was reached (3-4 days), the cells were serially passaged a total of six times. SK-N-SH cells were purchased from the National Biomedical experimental cell resource bank (Beijing, China), and were cultured in
Dulbecco's modi ed Eagle medium containing 10%
fetal bovine serum (Gibco, USA) at 37°C in a 5% CO 2 atmosphere.
Chang S., Min J., Lü X., Zhang Q., Shaofang S., Zhang T., & Wang L. (2022). Effect of epigenetic activating of Dlk1-Dio3 imprinted cluster on miR-370 expression due to folate deficiency during nerve development.
