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Moreover, donor-derived stem cells could be isolated from primary recipients, cultured as secondary spheres and after re-transplantation ameliorate radiation damage.
These were reported to be isolated from primary prostatic tumours.
Encouraged by the findings described above, we investigated whether similar stem cell precursors could be isolated from primary fibroblast cultures derived from subjects with HGPS.
The low number of cancer stem cells that can be isolated from primary tumors also limit the scope of metabolic study in this subpopulation of cells.
The existence of CSCs in solid tumours was first demonstrated in breast cancer in 2003; since then other studies have also shown that a CSC population can be isolated from primary breast tumours [ 1- 3].
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Single cells were isolated from primary tumors using the same procedure employed for MEC isolation.
Macrophages, the target cell for CSFV infection, were isolated from primary cultures of peripheral blood mononuclear cells, allowing us to utilize identical uninfected macrophages at the same time points as CSFV-infected macrophages, allowing only genes induced by CSFV to be identified.
Neurons, astrocytes and microglia were isolated from primary fetal human cortical cultures as described previously [16].
Total RNA was isolated from primary tumor cells and used for the synthesis of cDNA.
Genomic DNA was isolated from primary MM cell cultures at passages ≤6.
RNA was isolated from primary keratinocytes using Qiagen RNeasy Isolation Kit followed by cDNA synthesis (SuperScript II, Invitrogen).
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