e*******r
发帖数: 275 | 1 解剖打散之后出来的都是各种细胞的混合物吧, 怎么分离和处理才能拿到纯的神经元或
者纯的胶质细胞呢? 加arac是抑制astrocyte分裂的,不能杀死它们吧. 加不加BSA会不
会有不一样? | h*******o
发帖数: 4884 | 2 For primary neurons, we use neurobasal media +B 27. It is selective towards
neurons and glia usually cannot survive for long term in NBM+B27. With that
said, there is no pure neuronal culture. Anyway, cultured neurons do exhibit
some glial properties, such as glucose transporter expression, metabolic
activity, etc.
For embryonic glial culture, we use DMEM/F12 (1:1) +10% FBS. Because glial
cells are mitotic whereas neurons are post-mitotic, neurons will be easily
outnumbered by proliferative glia within days.
If you want enriched astrocytes, shake your mixed glial culture vigorously
for 2 to 4 hours to shake off microglia.
Always keep in mind, there is NO absolutely PURE neuronal or astrocytical
cultures.
Hope that helps. | e*******r
发帖数: 275 | 3 thanks so much
towards
that
exhibit
【在 h*******o 的大作中提到】
: For primary neurons, we use neurobasal media +B 27. It is selective towards
: neurons and glia usually cannot survive for long term in NBM+B27. With that
: said, there is no pure neuronal culture. Anyway, cultured neurons do exhibit
: some glial properties, such as glucose transporter expression, metabolic
: activity, etc.
: For embryonic glial culture, we use DMEM/F12 (1:1) +10% FBS. Because glial
: cells are mitotic whereas neurons are post-mitotic, neurons will be easily
: outnumbered by proliferative glia within days.
: If you want enriched astrocytes, shake your mixed glial culture vigorously
: for 2 to 4 hours to shake off microglia.
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