R28 cell line细胞株细胞系 BioVector NTCC质粒载体菌种细胞基因保藏中心

R28 cell line细胞株细胞系 BioVector NTCC质粒载体菌种细胞基因保藏中心
The R28 cells were developed from E1A-NR.3 parental cell line through three rounds of limiting dilution and were therefore derived from a single cell. Despite their clonal origin, these cells display both glial and neuronal cell markers indicative of a retinal precursor cell. The parental line E1A-NR.3 was established by immortalization of postnatal day 6 rat neuroretinal tissue using the psi2 replication incompetent retroviral vector. As a result, these cells are already resistant to geneticin/G418 and would require an alternative selection marker for transfection studies. These cells were designed not to form tumors in animals. Product Type: Cell Line Name: R28 Cell Type: 12S E1A-immmortalized rat retinal cells Organism: Rat; NOTE: The R28 cells have undergone verification by IDEXX-RADIL to be of rat origin without contamination by other mammalian cell lines. A baseline genetic profile of these cells is available upon request. Source: Postnatal day 6 rat neural retina Morphology: Adherent with glial and neuronal morphologies Biosafety Level: BSL-1 Subculturing: Split 1:2 when 80% confluent Growth Conditions: DMEM with 10% calf serum Cryopreservation: 90% culture medium with 10% DMSO

Care of the R28 retinal precursor cell line Gail M. Seigel, Ph.D. Thawing cells: Place a vial of cells in 37º water bath until thawed. Slowly pipet the cells, under sterile conditions, into a 10 mm dish or a T75 flask in DMEM with 10% serum (see recipe below). As soon as the cells are attached, remove the medium and add fresh medium to the dish. This gets rid of the DMSO that was in the original freezing vial. Ordinarily, every 2-3 days the cells either need to be fed or trypsinized. Trypsinization is accomplished as follows: Rinse cell surface with 1 ml CMF-EDTA, pour off, then add 1 ml CMF-EDTA. Add 1 ml of 0.125% trypsin to the dish and incubate at 37ºC for 3-5 min. Pipet the trypsin/CMF-EDTA solution over the cell sheet, mix well, and transfer a portion of the cell suspension to a fresh dish with DMEM+. I recommend a 1:2 or 1:3 split. If the cells get too sparse or too dense, they tend to die. However, there is a rather large window of acceptable cell density. I would recommend scaling up the cell culture and freezing down aliquots for future use. For freezing, we centrifuge cells at low speed, resuspend the cell pellet in 900 µl of DMEM+, and add 100 µl of Dimethyl Sulfoxide. Prefreeze at -70ºC overnight, then transfer to liquid Nitrogen. R28 cell description: R28 is an adherent retinal precursor cell line derived from postnatal day 6 Sprague-Dawley rat retina immortalized with the 12S E1A gene of adenovirus. The 12S E1A gene was introduced via an incompetent retroviral vector; therefore, no infectious virus is produced by R28 cells. The cells have been passaged 200 times thus far, and show no signs of senescence.
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