Gibco® Human Astrocytes and Gibco® Astrocyte Medium
Gibco® Human Astrocytes are normal cells derived from human brain tissue. When used with Gibco® Astrocyte Medium, Gibco® Human Astrocytes give you homogenous matured cells with proper large cell size and phenotype marker of GFAP. Due to their high biological relevance, Gibco® Human Astrocytes are ideal for studies of fundamental human neurological pathways, such as those involved in neurodegenerative disorders (e.g., Parkinson’s and Alzheimer’s diseases) as well as for neural injury studies (e.g., stroke).
Gibco® Human Astrocytes
Gibco® Human Astrocytes are human brain progenitor-derived astrocytes that are supplied cryopreserved at a concentration of ≥1 × 106 cells/mL in Gibco® Astrocyte Medium without EGF and with 10% DMSO. They exhibit ≥70% viability after thawing and stain positive for the astrocyte-specific marker glial fibrillary acid protein (GFAP).
Gibco® Astrocyte Medium
Gibco® Astrocyte Medium (sold separately or as part of a kit with cells) has been specifically formulated for the growth and maintenance of human and rat astrocytes while retaining their phenotype. The medium has three components: basal medium (DMEM), N-2 Supplement, and One Shot™ Fetal Bovine Serum (FBS). Epidermal growth factor (EGF) may also be added to enhance astrocyte proliferation.
Gibco® Human Astrocytes in Gibco® Astrocyte Medium
Figure 1. Gibco® Human Astrocytes can be recovered and maintained for 14 days retaining their phenotype of GFAP expression. (A) Phase contrast image. (B) GFAP expression (red).
Examples of cellular assays using human astrocytes
Using Gibco® Human Astrocytes and BacMam-delivered reporter gene constructs, experiments to measure calcium influx in response to agonists (Figure 2) and p53 activity (Figure 3) are conducted in efficient, easy-to-optimize workflows.
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Figure 2. Analysis of intracellular calcium flux in Gibco® Human Astrocytes using the Fluo-4 NW Calcium Assay Kit and BacMam Aequorin.* Gibco® Human Astrocytes were loaded with Fluo-4 NW (Cat. No. F36206) and tested for response to agonists. Emission values at 530 nm were obtained using the Hamamatsu Functional Drug Screening System (FDSS7000), and the relative fluorescence (in RFU) was plotted against the indicated compounds or different concentrations of L-quisqualic acid. Both (A) single-dose results and (B) the dose response curve revealed a significant response from the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) and group I metabotropic glutamate receptor agonist L-quisqualic acid. Gibco® Human Astrocytes transduced with BacMam Aequorin were treated with the agonist 48 hours post-transduction. The luminescence readout was obtained using the FDSS7000, and the relative luminescence (in RLU) were plotted against the indicated compounds or different concentrations of L-quisqualic acid. Both (C) single-concentration results and (D) the dose response curve revealed a significant response from the AMPAR and group I metabotropic glutamate receptor agonist L-quisqualic acid. [Note: *BacMam Aequorin materials are not catalog products. For more information, please contact us at firstname.lastname@example.org.]
Figure 3. BacMam transduction and Invitrogen™ BacMam-enabled cellular assays. (A) Human astrocytes transduced with 2.5% v2 CMV-GFP BacMam overnight revealed robust GFP expression after 48 hours in GFAP-positive cells. Primary antibody: rabbit anti-GFAP (Cat. No. 18-0063; 1:200 dilution); secondary antibody: goat anti–rabbit IgG (Cat. No. A11008; 1:500 dilution). (B) p53 posttranslational modifications in stressed cells. Invitrogen™ BacMam p53 [pSer15] Cellular Assay Kit (Cat. No. A12902) in Gibco® Human Astrocytes. Astrocytes were transduced with 10% v2 GFP-p53 virus directly in 384-well plates seeded with 1 x 104 cells/well. Data represent an etoposide (genotoxic damaging agent) serial dilution from 40 µM.