Gibco Rat Primary Cortical Astrocytes
Gibco Rat Primary Astrocytes exhibit superior cell viability and morphology and improved lot-to-lot consistency, providing neuroscience researchers an ideal, ready-to-use substitute to freshly isolated astrocytes. Upon expansion, more than 80% of the cells express the astrocyte-specific marker glial fibrillary acidic protein (GFAP).
Gibco Rat Primary Astrocytes
Gibco Rat Primary Cortical Astrocytes are isolated from the cortices of Sprague-Dawley rats at embryonic day 19 (E19) and cryopreserved at the end of the first passage (the cells can be further expanded for at least two additional passages). Each vial contains 1 × 106 cells. They exhibit ≥70% viability after thawing, and ≥80% stain positive for the astrocyte-specific marker glial fibrillary acid protein (GFAP).
Gibco Rat Primary Cortical Astrocytes give consistent post-thaw viability in the range of 70–90%, while a competitor’s cell viability was only 50% post-thaw (Figure 1).
Figure 1. Viability was determined by trypan blue exclusion using the Countess Cell Counter. Gibco Rat Primary Cortical Astrocyte viability is more than 80%, while astrocyte viability from a competitor is about 50%.
Gibco Rat Primary Cortical Astrocytes are cryopreserved at the end of passage 1 (P1), and can be recovered and further expanded for at least 2 additional passages (Figure 2).
Figure 2. Rat Cortical Astrocytes were thawed and cultured in astrocyte medium containing 85% high glucose DMEM and 15% fetal bovine serum. Cells were passaged every eight days by dissociation and replating on a plastic culture vessel.
Gibco Rat Primary Cortical Astrocytes stain positive for astrocyte-specific marker glial fibrillary acidic protein (GFAP) upon expansion. Figure 3 shows more than 80% of cells are positive for GFAP and less than 10% of the cells stain positive for neuron and oligodendrocyte markers.
Figure 3. Fluorescence images (20X) of Rat Cortical Astrocytes at P2 cultured for 3 days and stained for the appropriate phenotypic markers. Nuclei were stained with DAPI (blue). While approximately 80% of the cells stain positive for the astrocyte marker GFAP (A), less than 10% of the cells are positive for immature neuronal marker doublecortin (DCX) (B) and oligodentrocyte marker galactosylceramide (GalC) (C).
Q. From what age and strain of rat are Gibco Rat primary cortical astrocytes derived?
A. Gibco Rat primary cortical astrocytes are isolated from the cortex of fetal (E19) Sprague Dawley rats.
Q. At what passage were Gibco Rat primary cortical astrocytes cryopreserved?
A. Gibco Rat primary cortical astrocytes were cryopreserved in growth medium containing 10% DMSO at passage 1 (P1).
Q. What is the difference, if any, between cortical and hippocampal astrocytes?
A. There are few known differences between cortical and hippocampal astrocytes. However, it has been reported that astrocytes from different regions of the brain show a differential sensitivity to ischemic injury (Zhao and Flavin, 2000; Xu et al. 2001).
Q. Is there any difference between cortical astrocytes isolated from fetal, postnatal and adult tissue?
A. Primary rat astrocytes are typically isolated from late fetal or early postnatal brain because the resulting astrocytes are more viable in culture and of higher purity than astrocytes from adult brain.
Q. What is the viability of the cells once thawed?
A. Upon thawing, the vial has more than 70% viability, which will give more than 1 million live cells.
Q. What is the % purity of these cells?
A. Cell purity was evaluated in terms of the expression of astrocyte specific maker GFAP. More than 80% of the cells were positive for GFAP marker and the expression of neuronal marker (DCX) and oligodendrocyte marker (GalC) were less than 10%.
Q. How many passages can these cells be expanded?
A. Gibco Rat primary cortical astrocytes can be thawed, recovered and proliferated up to three passages to yield 1.5–2 fold increase of cells.
Q. What is the doubling time of the cells?
A. The doubling time of Gibco Rat primary cortical astrocytes is about 8.7 days.
Q. How can the cells be coated on glass coverslips?
A. Clean the coverslips and then coat them with 0.01% poly-D-lysine in double distilled water.
Q. I used astrocytes from another company and observed cell aggregates after thawing but not the astrocytes from your company. Why?
A. Gibco Rat primary cortical astrocytes were dissociated into single cell before cryopreservation, which gives an advantage to control the number of cells to plate.
Q. What are some of the key applications for astrocytes?
A. Astrocytes can be used to research cell transplantation, intracellular transport, ion-channel function, toxicity, receptor signaling, analyses of electophysiological properties, neurotransmitter release and evaluation of interaction between neurons and astrocytes.
Q. What related products can be used with these cells?
Key references for Gibco Rat Primary Cortical Astrocytes
- Pekny M, and Nilsson M (2005) Astrocyte activation and reactive gliosis. Glia 50:427–434.
- Rothstein JD, Martin L, Levey AI et al. (1994) Localization of neuronal and glial glutamate transporters. Neuron 13:713–725.
- Rothstein JD, Dykes-Hoberg M, Pardo CA et al. (1996) Knockout of glutamate transporters reveals a major role for astroglial transport in excitotoxicity and clearance of glutamate. Neuron 16:675–686.
- Lepore AC, Rauck B, Dejea C et al. (2008) Focal transplantation-based astrocyte replacement is neuroprotective in a model of motor neuron disease. Nat Neurosci 11:1294–1301.
- Maragakis NJ, Rothstein JD (2006) Mechanisms of Disease: astrocytes in neurodegenerative disease. Nat Clin Pract Neurol 2:679–689.
- Takano T, Oberheim N, Cotrina ML et al. (2009) Astrocytes and ischemic injury. Stroke 40:S8–12.