R28 Cell Line

Brief Introduction
Product List

General Information
Organism	Rattus norvegicus, rat
Cell Line Description	R28 is a conditionally immortalized rat retinal precursor cell line established by Dr. Gail M. Seigel in 1994 (and extensively characterized in 1996). This cell line was isolated from the intact neural retinal tissue of a female Sprague-Dawley rat at postnatal day 6. Its immortalization was achieved through transduction with a replication-defective ψ2 retroviral vector carrying the Adenovirus 12S E1A gene. The name "R28" both denotes its origin ("R" for retina) and indicates that it was selected from the 28th well of a 96-well plate during a limiting dilution cloning experiment. The R28 cell line has since become a premier, globally utilized model for studying neural retinal development. Although R28 was cloned from a single progenitor line (E1A-NR.3), it retains a heterogeneous precursor cell phenotype with multipotent differentiation potential. This cell line is widely employed in research fields related to retinal cell behavior, neuroprotective pathways, mechanical and chemical cytotoxicity, and the differentiation of glial cells into neuronal lineages.
Tissue	Eye; Neuroretina
Cell Type	Retinal Precursor Cell
Disease	Non-tumor (Derived from normal postnatal eye tissue)
Morphology	Polygonal; adherent epithelial-like and spindle-like heterogeneous mixture
Gender	Female
Age	Postnatal Day 6 (P6)
Product Format	Frozen
Growth Mode	Adherent (Anchorage-dependent)
Biosafety Level	2 (Sourced from mammalian tissue and immortalized using viral gene machinery; handled at BSL-2 globally to minimize potential biological exposure risks)
Applications	1. High-throughput in vitro screening of retinal neuroprotectants and antioxidants; 2. Elucidation of the mechanisms underlying light-induced phototoxicity and chemical cytotoxicity within the retinal neurosensory layer; 3. Construction of cellular mechanistic models for stress factors associated with diabetic retinopathy and glaucoma; 4. Investigation of the mechanisms governing transcriptional transitions and lineage specification during the differentiation of retinal progenitor cells into glial cells or functional photoreceptors; 5. Assessment of target gene expression levels using transcriptome microarrays or functional siRNA knockdown experiments.
Shipped In	Dry ice
Storage Temperature	−196°C (Liquid nitrogen vapor phase only)
Characteristics
Tumorigenic	No, exhibits contact inhibition and non-tumorigenic properties in standard tissue layouts
Lineage Relationship	Clonal child derivative obtained via three sequential rounds of limiting dilution from the parental E1A-NR.3 cell line (CVCL_5I34).
Transformant	Adenovirus 12S E1A integrated via replication-defective ψ2 retrovirus.
Growth Kinetics	Rapid adherent proliferation; typical population doubling time is 24 hours under optimal physical placement.
Mycoplasma Test	Negative
Culture Conditions and Handling
Subculturing	1. Remove and discard the spent complete medium from the culture dish. 2. Gently rinse the cell monolayer with standard Ca²⁺/Mg²⁺-free Dulbecco’s PBS (DPBS) or an EDTA solution to remove residual serum components containing active trypsin inhibitors. 3. Add 2.0 to 3.0 mL of pre-warmed 0.125% to 0.25% Trypsin-0.53 mM EDTA solution to completely cover the cell layer. 4. Incubate at 37°C for 2 to 10 minutes; observe closely under an inverted microscope until the cells detach, round up, and slide cleanly off the surface of the culture dish. 5. Immediately add 5.0 mL of pre-warmed complete growth medium to the dish to thoroughly neutralize the trypsin activity. 6. Centrifuge the suspension at 125 × g for 5 to 7 minutes; carefully aspirate the supernatant, uniformly resuspend the cell pellet in fresh medium, and aliquot into new culture vessels.
Medium Renewal	2 to 3 times per week
Subcultivation Ratio	Split confluent cultures at a standard ratio of 1:3 to 1:6.
Culture Conditions	Atmosphere: Air, 95%; CO2, 5%; Temperature: 37°C
Cryopreservation	90% Complete growth medium (with non-heat-inactivated FBS) + 10% DMSO

Cat.No.	Product Name	Price
CSC-RO02618	Cas9 Stable Cell Line - R28	Inquiry
CSC-RR00909	GFP Reporter Cell Line - R28	Inquiry
CSC-RR00968	Luciferase Reporter Cell Line - R28	Inquiry

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* For research use only. Not intended for any clinical use.

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