Cell line selection and supplementation

1. BasicsandDefinitions Cell lineselectionandsupplementation CEAC514 Assoc. Prof. Dr. Yasemin G. İşgör

2. Cell typeselection • The cell type chosen will depend on the question being asked. • DNA synthesis, • Response to cytotoxins, • Apoptosis, • Monitoringtheproduction of factors, proteins, enzymes, • Toxicitytestingorpharmacueticalassaystoevaluateeffects on organelles • Cell lineselectionbeginswiththefirst step as literaturesurvey • to determine from the literature whether a cell line exists with therequired properties. • Thecelllineto be studiedanditspossiblecontrols as cellareavailableor not • Theresearchorstudy plan specificsandassayconditioncompatibilitywithcellanditssupplements • The cell types may not matterprovided the cells are competentandassayconditionstoperform a researchactivity is compatiblewithculturingconditionsor can be controlledeasily.

3. Cell typeselection • Normal andhealthycellsortumorigeniccellsarethe main objectivetoperform a TCC task • Normal tissuecellshavefinite lifespan, while culturesfrom tumoursgiverisetocontinuous cell lines. • However, there are several • examples of continuous cell linesderived from normaltissues and which are non-tumorigenic. • Somerepresentativecells of thistypeare: • BHK-21 hamster kidney fibroblasts, • MDCKdog kidney epithelium, • 3T3 fibroblasts.

4. Cultureselectionbased on culturingandmaintenanceconditions • adeherentcellcultures • Culturesare propagated as a monolayer attached to the substratearecalledadeherentcellcultureswhichmay not needcentrifugationalprocedurewhileanalyzingtheculturemedia, orharvestingcellsformicroscopicexaminations, etc. • Adherentcellsareeasytohandleinterms of assayconditions but hard topassage, countandrevivecomparingtheprocedureswiththat of suspensioncells. • Suspension culture • Some of theadherentcellsortransformed cells, haematopoietic cells, and cells from ascitescan be propagated in suspension. • Suspension culture has advantages, includingsimpler propagation (subculture only requires dilution, no trypsinization), norequirement for increasing surface area with increasing bulk, ease of harvesting,andthe possibility of achieving a 'steady state' or biostat culture if required

5. Common Cell linesareused in TCC-1

6. Common Cell linesareused in TCC-2

7. Selection of Supplemetntarymediacompatiblewithcelllinesandculturingspecifics • Unfavorably the common attitude for medium and other supplement selection is based on 2 basic empirical rules: • Used formulations by others for the same cells, • Supplements availability: supplements are currently being used in the laboratory for different cells, • For continuous cell lines the formulation of supplements may not matter as long as the previously tested formulations providing the conditions as consistent. • For specialized cell types, for cells with genetic manipulations, and also for primary cells, favorable attitude for culture supplement selection is based on literature survey and cell line supplier’s MSDS and product data sheet suggestions. • If several media have been suggested in literature and bysupplierorresearchers in thesame lab, test them all, such as: • Measure the growth (population doubling time (PDT) • saturation density • expression of specific properties (differentiation,transfection efficiency, cell products, etc.). • Be awarethat the choice of medium maynot be the same in each case, • sowhenyou test eachorany of thesepleasekeep in mindthatthepreviousculturingconditions, taken as reference, must be thesimilarorthesame.

8. Commonlyusedmediaforcellcultures as part of supplementformulations Ref: Animal cell culture : a practical approach / edited byJohn R. W. Masters.—3rd ed., p.27