Oct 06, 2019
Preparation of primary chicken embryo liver (CEL) cells
- Norfitriah Mohamed Sohaimi1,
- Mohd Hair Bejo1,
- Abdul Rahman Omar1,
- Aini Ideris1,
- Nurulfiza Mat Isa2
- 1Faculty of Veterinary Medicine, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.;
- 2Faculty of Biotehnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
External link: https://doi.org/10.1371/journal.pone.0225863
Protocol Citation: Norfitriah Mohamed Sohaimi, Mohd Hair Bejo, Abdul Rahman Omar, Aini Ideris, Nurulfiza Mat Isa 2019. Preparation of primary chicken embryo liver (CEL) cells. protocols.io https://dx.doi.org/10.17504/protocols.io.7zchp2w
Manuscript citation:
Sohaimi NM, Bejo MH, Omar AR, Ideris A, Isa NM (2019) Molecular characterization of fowl adenovirus isolate of Malaysia attenuated in chicken embryo liver cells and its pathogenicity and immunogenicity in chickens. PLoS ONE 14(12): e0225863. doi: 10.1371/journal.pone.0225863
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 06, 2019
Last Modified: October 06, 2019
Protocol Integer ID: 28420
Materials
MATERIALS
Dulbecco’s Modified Eagle’s Medium (DMEM)Merck MilliporeSigma (Sigma-Aldrich)Catalog #D5796
Fetal Bovine SerumGibco - Thermo Fisher ScientificCatalog #10270106
Gibco Penicillin-Streptomycin (10,000 U/mL) (Pen/Strep)Fisher ScientificCatalog # 15-140-122
0.1M Phosphate Buffered Saline pH 7.4
Trypsin-EDTA (0.25%), phenol redThermo FisherCatalog #25200056
Primary CEL cell was obtained from liver embryos of 13 to 15 days old SPF embryonated chicken eggs. Liver was harvested aseptically using sterile forceps and washed twice with sterile phosphate buffered saline (PBS, pH 7.4, 0.1M). The liver tissue was minced and trypsinized gently with 0.25% Trypsin-EDTA solution for 10 minutes. The suspension was passed through muslin cloth and centrifuged at 96 x g for 5 minutes to obtain cell pellet. Trypsin was discarded and cell pellet was resuspended with fresh Dulbecco`s Modification Eagle Medium (DMEM), enriched with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin antibiotic. Cell concentration was counted and adjusted to 5 x106 cells/mL. The cell suspension (5mL) was seeded into new 25cm2 cell culture flasks and was kept under controlled atmosphere at 5% CO2 incubator with 85%-90% humidity until confluent monolayer formed [Soumyalekshmi et al., 2014].