From: Kinetic oxygen measurements by CVC96 in L-929 cell cultures
Cell line: | L-929 |
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Cell type: | mouse connective tissue fibroblast |
Origin: | established from the normal subcutaneous areola and adipose tissue of a male C3H/An mouse; used as target in TNF detection assays |
Morphology: | fibroblasts growing as monolayer |
Medium: | 90% RPMI 1640 + 10% FBS |
Subculture: | split confluent cultures 1:5 to 1:10 using trypsin (do not use trypsin/EDTA); after 2–3 days monolayer will be confluent; split 2–3 times a week; seed out initially at about 1.0–2.0 × 106 cells/25 cm2 |
Incubation: | at 37°C with 5% CO2 |
Doubling time: | ca. 21–24 hours |
Harvest: | ca. 4–8 × 106 cells/25 cm2 |
Storage: | frozen with 70% medium, 20% FBS, 10% DMSO at about 1–2 × 106 cells/ampoule |
Mycoplasma: | negative in DAPI, microbiological culture, PCR assays |
Species: | confirmed as mouse with IEF of AST, MDH, PEP B |
Cytogenetics: | murine hypertriploid karyotype – 61–67, 12–16 centric fusion markers present |
Viruses: | ELISA: reverse transcriptase negative |