PC12 rat pheochromocytoma cells were originally obtained from the American Type Culture Collection (CRL-1721; ATCC, Manassas, VA). MAEC (mouse aortic endothelial cell line), MVEC (mouse venous endothelial cell line) and HUVEC (human umbilical venous endothelial cell line; used as an “endothelial cell control”) in our studies were gifts from Dr. Dimitri Azar of University of Illinois Chicago [13 (link)]. Induced PC12 cells were grown on 100mm tissue culture petridishes pre-coated with collagen substrate, Collagen I solution (0.5 mg/mL, BD Biosciences) and maintained in RPMI-1640 medium supplemented with 10% heat-inactivated horse serum and 5% fetal bovine serum (FBS), 100 U/ml penicillin G, and 100 μg/ml streptomycin (Gibco, Grand Island, NY) and 50 ng/ml purified recombinant Mouse beta-NGF (R&D Systems) at 37°C under an atmosphere of 5% CO2 and 95% air. HUVEC, MAEC and MVEC were maintained in VascuLife® Endothelial Medium (containing EnGS (containing Endothelial Cell Growth Supplement; Lifeline® Cell Technology, Frederick, MD) at 37°C under an atmosphere of 5% CO2 and 95% air.
Collagen 1 solution
Manufactured by BD
Collagen I solution is a laboratory product that provides a concentrated source of purified type I collagen. It is a transparent, viscous liquid that can be used as a substrate for cell culture and various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using collagen 1 solution
1
Culturing Rat Pheochromocytoma and Endothelial Cells
2
Isolation of Whole Dorsal Root Ganglia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All primary cell isolation procedures were performed with the approval of the IACUC at the University of Florida and in accordance with guidelines established by the NIH.
Whole DRGs were obtained from P1-P3 mice. Briefly, using aseptic techniques, the spinal column was first removed. Next, using a stereomicroscope whole DRGs were pulled out from between each of the spinal segments (from cervical to lumber). Whole DRGs were stored and trimmed in 1x HBSS supplemented with 1% gentamicine on ice until ready for use. Isolated and trimmed DRGs were carefully picked up using microtweezer (Dumont #5) and dragged through a drop of collagen I solution (3mg/mL, BD Sciences, San Jose, CA). DRG were then placed on the substrates and cultured in neurobasal medium supplemented with 2% B27, 20 nm L-glutamine, 1% penicillin/streptomycin/antimicotic, and 50 ng/ml of nerve growth factor (NGF; R&D Systems, Minneapolis, MN) at 37°C for 3 days in vitro.
Whole DRGs were obtained from P1-P3 mice. Briefly, using aseptic techniques, the spinal column was first removed. Next, using a stereomicroscope whole DRGs were pulled out from between each of the spinal segments (from cervical to lumber). Whole DRGs were stored and trimmed in 1x HBSS supplemented with 1% gentamicine on ice until ready for use. Isolated and trimmed DRGs were carefully picked up using microtweezer (Dumont #5) and dragged through a drop of collagen I solution (3mg/mL, BD Sciences, San Jose, CA). DRG were then placed on the substrates and cultured in neurobasal medium supplemented with 2% B27, 20 nm L-glutamine, 1% penicillin/streptomycin/antimicotic, and 50 ng/ml of nerve growth factor (NGF; R&D Systems, Minneapolis, MN) at 37°C for 3 days in vitro.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!