RF/6A

RF/6A is a rhesus macaque (Macaca mulatta) retinal choroidal endothelial cell line established from fetal choroid and retina tissue. The line is grows as an adherent monolayer with epithelial-like morphology. RF/6A cells retain key endothelial characteristics including expression of Factor VIII (von Willebrand factor), fibronectin, and Weibel-Palade granules detectable by electron microscopy — the latter confirming their endothelial identity. The line was originally established for studies of retinal and choroidal vascularization and has been widely adopted as a primate endothelial model for ocular angiogenesis research.

RF/6A is applicable in ocular angiogenesis research, studies of retinal and choroidal vascularization, evaluation of anti-angiogenic agents (VEGF inhibitors, bevacizumab, ranibizumab), age-related macular degeneration (AMD) modeling, diabetic retinopathy biology, and assessment of vascular permeability in the ocular microenvironment. The non-human primate (NHP) origin makes RF/6A closer to human retinal vascular biology than rodent endothelial models, particularly for studies involving primate-specific VEGF isoform responses and ocular pharmacology. The line is commonly used in tube formation assays, migration assays, and VEGF-stimulation experiments.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.