Viral Manipulation of Neural Stem/Precursor Cells - Viral Vector Approaches in Neurobiology and Brain Diseases - page 271

Fig. 2 The viral particles obtained as shown in Fig. 1 can be used in different
contexts: to infect (a) neural stem cells isolated from the SVZ or (b) organotypic
slice cultures in vitro or to (c) directly infect cells in vivo, after viral particle injec-
tion into living animals

4 Notes

1. Incubation times may vary. The tissue pieces usually easily go
into solution by gentle pipetting.
2. In the first passages of culturing and enrichment of proliferat-
ing cells, the emerging neurospheres are mechanically dissoci-
ated until the preparation is cleaned from dissection debris and
the NPC line is stably growing. Afterwards the passaging is
performed enzimatically using Accumax.
3. Cell density for normal culturing and expansion of mNPCs are
8,000 cells/cm 2 . However, the plating density might vary
between different brain regions and applications [ 86 - 88 ].
4. Calcium phosphate precipitation is one of the most common
methods for transfecting packaging cells for lentivirus produc-
tion, providing high transfection efficiency while still having
extremely cheap costs. Other methods are commercially avail-
able and guarantee high efficiency transfection. These include
both liposomal and nonliposomal methods.