Cell line selection and supplementation Basics and Definitions CEAC514 Assoc. Prof. Dr. Yasemin G. İşgör

The cell type chosen will depend on the question being asked. Cell type selection The cell type chosen will depend on the question being asked. DNA synthesis, Response to cytotoxins, Apoptosis, Monitoring the production of factors, proteins, enzymes, Toxicity testing or pharmacuetical assays to evaluate effects on organelles Cell line selection begins with the first step as literature survey to determine from the literature whether a cell line exists with the required properties. The cell line to be studied and its possible controls as cell are available or not The research or study plan specifics and assay condition compatibility with cell and its supplements The cell types may not matter provided the cells are competent and assay conditions to perform a research activity is compatible with culturing conditions or can be controlled easily.

Some representative cells of this type are: Cell type selection Normal and healthy cells or tumorigenic cells are the main objective to perform a TCC task Normal tissue cells have finite lifespan, while cultures from tumours give rise to continuous cell lines. However, there are several examples of continuous cell lines derived from normal tissues and which are non-tumorigenic. Some representative cells of this type are: BHK-21 hamster kidney fibroblasts, MDCK dog kidney epithelium, 3T3 fibroblasts.

Culture selection based on culturing and maintenance conditions adeherent cell cultures Cultures are propagated as a monolayer attached to the substrate are called adeherent cell cultures which may not need centrifugational procedure while analyzing the culture media, or harvesting cells for microscopic examinations, etc. Adherent cells are easy to handle interms of assay conditions but hard to passage, count and revive comparing the procedures with that of suspension cells. Suspension culture Some of the adherent cells or transformed cells, haematopoietic cells, and cells from ascites can be propagated in suspension. Suspension culture has advantages, including simpler propagation (subculture only requires dilution, no trypsinization), no requirement for increasing surface area with increasing bulk, ease of harvesting,and the possibility of achieving a 'steady state' or biostat culture if required

Common Cell lines are used in TCC-1

Common Cell lines are used in TCC-2

Selection of Supplemetntary media compatible with cell lines and culturing specifics 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 by supplier or researchers in the same 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 aware that the choice of medium maynot be the same in each case, so when you test each or any of these please keep in mind that the previous culturing conditions, taken as reference, must be the similar or the same.

Commonly used media for cell cultures as part of supplement formulations Ref: Animal cell culture : a practical approach / edited by John R. W. Masters.—3rd ed., p.27