Expansion of Neuroprogenitor Cells

Neuroprogenitor cells were selectively expanded using non-adherent culture [16 (link),17 (link),18 (link),19 (link),20 ,21 (link)]. Briefly, aMSCs were seeded at 10,000 cells/cm² density onto Ultra Low^® non-adherent 6-well plates (Corning, Corning, NY, USA) in sphere-forming medium (SFM) comprising DMEM/F12 (Thermo Fisher Scientific) supplemented with 2% B27 (Thermo Fisher Scientific), bFGF, and EGF (20 ng/mL, Peprotech, Rehovot, Israel). Cultures were maintained for 8–10 days with medium refreshed every 48 h. At timed intervals, selected spheres were partially dissociated via incubation in TrypLE Express for efficient cell counting and determination of proportion of cells immunopositive for the neural stem/progenitor marker nestin [24 (link)] and the OL lineage determining factor Olig2 [25 (link),26 (link)].

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Tsui Y.P., Lam G., Wu K.L., Li M.T., Tam K.W., Shum D.K, & Chan Y.S. (2021). Derivation of Oligodendrocyte Precursors from Adult Bone Marrow Stromal Cells for Remyelination Therapy. Cells, 10(8), 2166.

Publication 2021

Ultra Low® non-adherent 6-well plates DMEM/F12 bFGF EGF

Cells Nestin Neural Olig2 Stem

Corresponding Organization : University of Hong Kong

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Variable analysis

independent variables

Culture method: non-adherent culture

dependent variables

Proportion of cells immunopositive for the neural stem/progenitor marker nestin
Proportion of cells immunopositive for the OL lineage determining factor Olig2

control variables

Cell seeding density: 10,000 cells/cm^2
Culture medium: DMEM/F12 supplemented with 2% B27, bFGF, and EGF (20 ng/mL)
Culture duration: 8-10 days
Medium refreshed every 48 h

positive controls

No positive controls explicitly mentioned

negative controls

No negative controls explicitly mentioned

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