- References (2)
- Inventor Info
|Antigen/Gene or Protein Targets
|Polycomb complex protein BMI-1, polycomb group RING finger protein 4; PCGF4; RING finger protein 51; RNF51; B cell-specific Moloney murine leukemia virus integration site 1; BMI-1; FLVI2/PCGF4; Mo-MLV; Moloney murine leukemia virus; NHBEC; Normal Human Bronchial Epithelial Cells; pLVX-BMI-1 Cell Line: COPD, ALI; air–liquid interface
Primary Human bronchial epithelial cells when grown in vitro have a limited lifespan and begin to deviate both in phenotype and morphology, losing the plasticity required around passage 4 or 5, for air-liquid interface (ALI) differentiation. These Human bronchial epithelial cells expressing BMI-1 retain both viability and differentiation potential of wild-type human bronchial epithelium while importantly not demonstrating changes in cell karyotype.
B lymphoma Moloney murine leukemia virus insertion region 1 homolog (BMI-1) is an oncogene which functions by regulating P16 and P19 cell cycle inhibitor genes and is also associated with erythroplakia and tongue cancer. BMI-1 is thought to repress, p16(Ink4a), a cyclin-dependent kinase inhibitor and tumor suppressor that induces cell cycle arrest at the Gap 1 phase. BMI-1 can therefore be used to delay cell senescence.
The airway epithelium is a critical interface acting as a barrier to potential pathogens and extraneous particles, assisting in regulation of host defense mechanisms like the inflammatory response.
|Passage 2 heterogeneous cells were plated in a 6-well plate at 5 x 10 4 cells per well and grown overnight. Media was replaced with 800 ul of media with 2 ug/ml of polybrene and 6.25 uL of lentivirus pLVX-BmI-1 to give >90% transfection. The plates were incubated at 37°C for 6.5 h with gentle rocking and the media replaced after 2 hours. After 72 h, selection media containing 300 ng/mL puromycin was added and cells with antibiotic resistance after 6 days of selection were considered to have been stably infected with virus expressing BMI-1.
|Cell Cycle, Cell Signaling & Signal Transduction, Cell Structure and Motility, Drug Discovery & Development, Genetic Studies Tools
|Recommended Growing Conditions
|Normal human bronchial epithelial cells were grown in a growth factor-supplemented medium and differentiated at ALI in bronchial epithelial differentiation medium.
Donor details: Male, 50 years old, Caucasian, smoker and consumed alcohol.
Cells were fixed for immunostaining, histology sectioning, and scanning electron microscopy after 28 days at ALI.
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