This video demonstrates the generation and differentiation of primary neurospheres. Neural stem cells from zebrafish, in the presence of growth factors, proliferate into free-floating neurospheres with self-renewal capacity, which later differentiate into glial cells and neurons.
Protocol
1. Preparations Prepare 10 ml of dissection medium, and add 200 µl of 100x penicillin-streptomycin into 9.8 ml of Dulbecco's Modified Eagle Medium F12 (DMEM/F12). Prepare L-Cysteine solution: to 10 ml of water for tissue culture, and add 120 mg of L-cysteine. Store the L-cysteine solution at 4 °C for up to 2 weeks or in aliquots at -20 °C for up to 1 year. Prepare DNase I solution: to 1 ml of water for tissue culture, add 10 mg of DNase I. Store the DNas…
Representative Results
Figure 1: Differentiation of Zebrafish Brain-derived Neurospheres. (A-C, E) Phase contrast images of whole brain-derived neurospheres cultured in the Z-differentiation medium during 1 (A, DiVd1), 3 (B, DiVd3) and 4 (C and D, DiVd4) days. (E) D…