Browser Not Supported.

IE users: please get the latest browser version at Microsoft Internet Explorer Home Page

if yot already have the latest browser version please verify that your browser settings is not in compatibility mode

Browser: unknown
Version: 0

All in Vitro Cells > Primary Cell card

Umbilical tissue-derived cells (CNTO2476) (Janssen Biotech, Inc)

Umbilical tissue-derived cells are isolated from minced and enzymatically digested umbilcal cords.

Umbilical cords are minced and enzymatically digested with Dulbecco's modified Eagle's low-glucose (Lg) medium (DMEM) mixed with collagenase (50 U/ml). The cell suspension is filtered through a 70-μm filter, and the supernatant is centrifuged at 350g. Isolated cells are washed in DMEM-Lg several times and plated at a density of 5,000 cells per cm2 in T75 flasks in DMEM-high glucose (Hg), supplemented with defined fetal bovine serum (FBS; 15%), nonessential amino acids (0.1%) , β-mercaptoethanol, penicillin (100 U/ml), and streptomycin (100 μg/ml). After cryopreservation, cells are thawed and seeded on gelatin-coated flasks, in growth medium (consisting of DMEM-Lg supplemented with FBS (15%) , β-mercaptoethanol ( 0.001%) , penicillin (100 U/ml) and streptomycin (100 μg/ml)). Cells are cultured under standard conditions in atmospheric oxygen with 5% carbon dioxide, at 37°C. When cells reach approximately 70% confluence, they are passaged every 3-4 days, using trypsin/EDTA, and reseeded at a density of 5,000 cells per cm2 onto gelatin-coated flasks. Flasks were gelatin coated by placing a solution of gelatin (2% (wt/vol)) into tissue culture flasks for a minimum of 20 minutes at room temperature. The solution is then aspirated, and the flasks are washed at least two times with phosphate-buffered saline (PBS) prior to cell culture use. Cells are harvested after 10 passages (approximately 20 population doublings) and cryopreserved in supplemented growth medium (Hg) containing DMSO (10%) in a programmable rate freezer.

See additional Stem, Progenitor & Primary Cells for: Umbilical Cord
Primary Cell
Homo sapiens
Umbilical tissue-derived cells (CNTO2476)