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Fig. 1. Single neural stem cells can differentiate into neurons, glia and myocytes.
A single retrovirally labeled E14 CD31- CD35- NSC was
expanded in suspension in MB-media for 6 days then transferred to an adherent
surface and allowed to differentiate. Differentiated progeny were identified
by immunostaining 14 days post-plating. (A-C) Epifluorescent images of an
expanding EGFP+ NSC. The resulting sphere was dissociated and grown
as an adherent culture for 14 days to complete the differentiation process.
(D) Presented are epifluorescent images of EGFP+ cells (green) and
sk-MHC immunoreactive cells (red). Arrowheads indicate a phenotypic
EGFP+ neuron in a cluster of EGFP+ sk-MHC+
myocytes. Scale bar: 80 µm. (E-H) The observed multilineage differentiation
covered many combinations of differentiation possibilities. (E,F) Confocal
z-sections of differentiated clones containing BAG5+
cardiac myocytes (red) and MAP2 immunoreactive neurons (green); (G,H) BAG5
cardiac myocytes (red) and GFAP+ glial cells (green). To further
verify that differentiated clones arose from a single EGFP+ cell,
Southern analysis was performed on random differentiated progeny; (I) Southern
blot. Clone 15-1 is the founder clone and clone 15-2 represents the secondary
clone arising from the primary clone 15-1. Controls 1 and 2 are mixtures of
two or more individual clones.
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