Neural stem cells (hsNSCs) were isolated from human fetal striatum and expanded in a serum- and growth factor-dependent adherent culture. hsNSCs are highly proliferative and multipotent cells, capable of differentiating into neurons, astrocytes, and oligodendrocytes. hsNSCs are morphologically homogeneous and express neural stem cell markers, but no neural differentiation markers.
Striatum tissue was obtained from miscarried human fetal samples at gestational age 12–20 weeks. The human fetal striatum tissue fragment was dissected, washed with PBS, mechanically cut into small pieces (~ 1 mm3), and triturated gently with a siliconized Pasteur glass pipette. Cell suspension was passed through a 70 µm strainer and cultivated in uncoated 75 cm2 cell culture flasks at a density of 1x106 cells/ml. The primary culture medium was Dulbecco's modified Eagle's medium/nutrient mixture F12 (DMEM/F12) supplemented with fetal bovine serum (10%), EGF (20 ng/ml), bFGF (20 ng/ml), and LIF (10 ng/ml). After 48–72 hours, the adherent clones were washed five times with fresh sterile PBS and replaced with fresh culture medium.
EMBRYONIC DEVELOPMENT & STEM CELL COMPENDIUM
