The presence of platelet-derived growth factor AA homodimer (PDGF-AA) and basic fibroblast growth factor (bFGF) in complete GPC growth medium allows rat GPCs to remain undifferentiated. Figure 1 shows more than 80% of the GPC population retain the undifferentiated phenotypic marker A2B5.
|Figure 1. Rat GPCs stained by indirect immunofluorescence for the cell-surface marker A2B5 (green). Nuclei were stained with DAPI (blue). Cells were maintained in the undifferentiated state in GPC recovery media for three days prior to 4% paraformaldehyde fixation and staining.|
Upon expansion, rat GPCs retain their differentiation potential into oligodendrocytes and astrocytes. Figure 2 shows rat GPCs differentiating into oligodendrocytes. More than 30% of rat GPCs were stained positive for GalC (oligodendrocyte specific marker).
|Figure 2. Rat GPCs differentiating to oligodendrocytes. The cells were differentiated in growth medium lacking PDGF-AA, EGF, and bFGF, but supplemented with 2% FBS for three days prior to fixation with 4% paraformaldehyde and subsequent staining. The cell-surface marker GalC was detected by indirect immunofluorescence (green) in ≥30% of the cells in the culture. Nuclei were stained with DAPI (blue).|
Gibco rGPCs can be further expanded for at least one passage upon thawing in GPC recovery media (StemPro NSC SFM, GlutaMAX-I, and PDGF-AA). The proliferation abilities afford researchers a more than 2-fold increase in cell numbers while the cells retain their typical morphology (Figure 3).
|Figure 3. Brightfield image of Rat GPCs at passage 3 (P3) that have been cultured in complete StemPro NSC SFM supplemented with 2 mM GlutaMAX-I and 10 ng/mL PDGF-AA (i.e., complete GPC growth medium) for three days. The image was captured using a 10X objective lens.|
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