Table 7.

Proliferation assays for cigarette smoke

AssayEnd point measuredCell studiedSmoke conditionsCigarette typeTest Substance*Proliferation results reported in studyReference
Attachment assayNo. of cells that detach after treatmentMouse embryonic fibroblasts, D3 mice embryonic stem cellsN/A2R1, 1R4F, and four commercial cigarettesMS (Solvent trap)Dose-dependent response observedLin, S 2009 (161)
BrdUrd incorporation using Nova REDCells counted that dyed redHuman pulmonary muco-epidermoid carcinoma cell line NCI-H292, primary bronchial epithelial cellsN/ACommercial cigarette and 1R3CSC (Solvent trap)Dose-dependent response observed at lower concentrations, proliferation decrease at higher concentrationsLuppi, F 2005 (73)
BrdUrd incorporation FACS analysisCells counted that incorporate BrdUrdT cells extracted from peripheral blood mononuclear cellsN/A1R4FCSE (Solvent trap)CSE treatment resulted in a significant reduction in proliferationGlader, P 2006 (75)
3H Thymidine uptakeCounting the number of cells that incorporate radioactive labelAlveolar type II epithelial cell line A549N/ACommercial medium tar cigaretteCSC (Solvent trap)Dose-dependent response observed at higher concentrationsLannan, S 1994 (150)
Human lung epithelial cell line A549N/AAn Indian cigarette with filterCSC (Solvent trap)Proliferation was increased at lower concentrations but was inhibited at higher concentrations.Kaushik, G 2008 (76)
MTTReduction of tetrazolium salts to formazan dyes by cellular enzymesHuman normal breast epithelial cell line MCF10AFTC1R4FTPM (Cambridge method)Concentration-dependent response observed.Narayan, S 2004 (21)
Human epidermal keratinocytes and oral carcinoma cellsFTC1R3FCSC (Cambridge method)Concentration- and time-dependent response observedNagaraj, NS 2006 (48)
Human lung epithelial cell line A549N/AAn Indian cigarette with filterCSC (Solvent trap)Concentration response observedKaushik, G 2008 (76)
Human pulmonary artery endothelial cellsN/ALong Life cigarettesCSE (Cambridge method)Dose-dependent decrease observedHsu, CL 2009 (159)
Human lung epithelial cells A549N/ACommercial filtered cigaretteAECS (Solvent trap)Dose- and time-dependent decrease observedDas, A 2009 (152)
Survival/Proliferation assayCount the number of cells per colony using a computerized image analyzerHuman bronchial epithelial cellsN/AN/ACSC fractionsDose-dependent inhibition of proliferation observedWilley, JC 1987 (23)
Survival/Proliferation assayCounting colonies and stained cellsWild-type and mutated CHO cell linesN/A1R4FTPM (Cambridge method)Concentration-dependent response observed differed by cell linesKato, T 2007 (52)
Survival/Proliferation assayCells counted by Coulter counterNormal human bronchial epithelial cellsFTC2R4F,Quest low nicotine and nicotine freeTPM (Cambridge method)Dose-dependent suppression of cell proliferation observedChen, J 2008 (146)
WST-1Reduction of tetrazolium salts to formazan dyes by cellular enzymesLung epithelial cellsISOA major international brandGVP (Exposure chamber)Dose-dependent response observedPiperi, C 2003 (92)
Mouse lung epithelial cell line LA-4 cellsISOCigarettes with different filtersGVP (Exposure chamber)Proliferation differed by filter typePouli, AE 2003 (94)
Human fetal bronchial epithelial cellsFTCResearch cigarette type A and BWS (Exposure chamber), smoke diluted with synthetic airDifferent effects by air dilution and cigarette typeRitter, D 2003 (85)
Human lung epithelial cell line A549ISO2R4F, 2R4F with modified acetate or charcoal filterWS and GVPViability decreased as exposure increased, dose-dependent response observedFukano, Y 2004 (83)
Human gingival fibroblastsN/AN/ACSCHigher concentrations decrease proliferationZhang, W 2009 (151)
WST-8Reduction of tetrazolium salts to formazan dyes by cellular enzymesNormal human bronchial epithelial cellsFTC2R4F,Quest low nicotine and nicotine freeTPM (Cambridge method)Dose-response observedChen, J 2008 (146)
XTT assayActivity of mitochondrial dehydrogenasesCHO WBL strainFTC1R4FTPM (Cambridge method)Proliferation observed after1-6 h of exposurePutnam, KP 2002 (44)

Abbreviations: FACS, fluorescence-activated cell sorting; HPAEC, human pulmonary artery endothelial cell.

  • *TPM or CSC indicated as reported in publication, but actual method shown in parenthesis.