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  /Title (ptc-c8-basic-cell-culture-medium.pdf)
  /Creator (Cheatography)
  /Author (woozing)
  /Subject (PTC - C8 (Basic Cell Culture Medium) Cheat Sheet)
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    \vspace{-2pt}\large{\bf{\textcolor{DarkBackground}{\textrm{PTC - C8 (Basic Cell Culture Medium) Cheat Sheet}}}} \\
    \normalsize{by \textcolor{DarkBackground}{woozing} via \textcolor{DarkBackground}{\uline{cheatography.com/146689/cs/31846/}}}
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  \mymulticolumn{2}{p{5.377cm}}{\bf\textcolor{white}{Cheatographer}}  \\
  \vspace{-2pt}woozing \\
  \uline{cheatography.com/woozing} \\
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  \mymulticolumn{1}{p{5.377cm}}{\bf\textcolor{white}{Cheat Sheet}}  \\
   \vspace{-2pt}Not Yet Published.\\
   Updated 26th April, 2022.\\
   Page {\thepage} of \pageref{LastPage}.
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  Measure your website readability!\\
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\begin{tabularx}{8.4cm}{x{2.584 cm} x{2.508 cm} x{2.508 cm} }
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\mymulticolumn{3}{x{8.4cm}}{\bf\textcolor{white}{Media}}  \tn
% Row 0
\SetRowColor{LightBackground}
1. Natural Media & a) Coagulant, such as plasma clots & -Blood before coagulation; Plasma can also be prepared in the laboratory by taking out blood and adding heparin to prevent blood coagulation. \tn 
% Row Count 11 (+ 11)
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\SetRowColor{white}
-natural sources of nutrient sufficient for growth and \seqsplit{proliferation} of animal cells and tissues & b) Biological fluids such as serum & -Blood after coagulation; serum: components of animal cell culture which is the source of various nutrients. \tn 
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 &  & -serum: Contains growth factors which promotes cell \seqsplit{proliferation}, cell attachment and adhesion factors \tn 
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 &  & -other forms of biological fluids used are coconut water, amniotic fluid, pleural fluid, insect haemolymph serum, culture filtrate, aqueous humour, from eyes etc. \tn 
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\begin{tabularx}{8.4cm}{x{2.584 cm} x{2.508 cm} x{2.508 cm} }
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\mymulticolumn{3}{x{8.4cm}}{\bf\textcolor{white}{Media (cont)}}  \tn
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 & c) Tissue extracts for example Embryo extracts & -Extracts from tissues such as embryo, liver, spleen, leukocytes, tumour, bone marrow etc. are also used for culture of animal cells. \tn 
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2. Synthetic Media & a) Serum containing media &  \tn 
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-prepared artificially & b) Serum-free media &  \tn 
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\mymulticolumn{3}{x{8.4cm}}{{\emph{-Recipe: organic and inorganic nutrients, vitamins, salts, serum proteins, carbohydrates, co-factors, etc. }}} \tn 
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\begin{tabularx}{8.4cm}{p{0.8 cm} p{0.8 cm} }
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Base Media}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-many different types of base medium available} \tn 
% Row Count 1 (+ 1)
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\mymulticolumn{2}{x{8.4cm}}{-Additional supplements are added to this bottle} \tn 
% Row Count 2 (+ 1)
% Row 2
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\mymulticolumn{2}{x{8.4cm}}{-Some cell types prefer one type of medium to another, be sure to know all this information ahead of time.} \tn 
% Row Count 5 (+ 3)
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\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Types of Base Medium}}  \tn
\SetRowColor{LightBackground}
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\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Basic Constituents of Media}}  \tn
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\SetRowColor{LightBackground}
Constituents & Function \tn 
% Row Count 1 (+ 1)
% Row 1
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1. Inorganic Salts & -retain the osmotic balance of the cells \tn 
% Row Count 3 (+ 2)
% Row 2
\SetRowColor{LightBackground}
 & -regulate membrane potential by provision of sodium, potassium and calcium ions \tn 
% Row Count 7 (+ 4)
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\SetRowColor{white}
 & -required in the cell matrix for cell attachment and as enzyme cofactors \tn 
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2. Carbohydrates (glucose) & -main source of energy \tn 
% Row Count 13 (+ 2)
% Row 5
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{\emph{major sugars used are glucose and galactose, however, some media contain maltose or fructose}} & -media containing the higher concentration of sugars are able to support the growth of a wider range of cell types \tn 
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\SetRowColor{LightBackground}
3. Amino Acids & -building blocks of proteins \tn 
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 & -'Essential' amino acids must be added to culture media as cells are not able to synthesize these themselves \tn 
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 & -The concentration of amino acids in the culture medium will determine the maximum cell density that can be achieved \tn 
% Row Count 33 (+ 6)
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Basic Constituents of Media (cont)}}  \tn
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 & -once depleted the cells will no longer be able to proliferate \tn 
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 & -once depleted the cells will no longer be able to proliferate \tn 
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 & -Commonly the necessary amino acids include cysteine and tyrosine, but some non-essential amino acids may be needed. \tn 
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 & -Adding supplements of non-essential amino acids to media both stimulates growth and prolongs the viability of the cells in culture. \tn 
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4. Vitamins & -precursors for numerous co-factors \tn 
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 & -Many vitamins, especially B group vitamins, are necessary for cell growth and proliferation and for some lines the presence of B12 is essential. \tn 
% Row Count 31 (+ 8)
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\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Basic Constituents of Media (cont)}}  \tn
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 & -Many vitamins, especially B group vitamins, are necessary for cell growth and proliferation and for some lines the presence of B12 is essential. \tn 
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5. Fatty Acids and Lipids & -important in serum free media since they are normally present in serum \tn 
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 & -E.g. cholesterol and steroids essential for specialised cells. \tn 
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6. Proteins and Peptides & -particularly important in serum free media \tn 
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 & -most common proteins and peptides include albumin, transferrin, fibronectin and fetuin and are used to replace those normally present through the addition of serum to the medium \tn 
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7.  Buffering Systems - pH & -Close control of pH is essential for optimum culture conditions \tn 
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\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Basic Constituents of Media (cont)}}  \tn
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 & -Most cells require pH conditions in the range 7.2-7.4 \tn 
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 & -Phenol red indicator to monitor pH (pH 6.5 = yellow, pH 7.0 = orange, pH 7.4 = red, pH 7.8 = purple) \tn 
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 & -Could monitor contamination \tn 
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8. Trace Elements & -E.g. zinc, copper, selenium and tricarboxylic acid intermediates \tn 
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% Row 25
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 & -Selenium is a detoxifier and helps remove oxygen free radicals \tn 
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\SetRowColor{white}
9. Antibiotics and Anti-mycotics (anti-fungal) & -Unless good sterile conditions can be maintained (e.g., using laminar flow hoods) it is necessary to incorporate antibiotics and antimycotics into the media. \tn 
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 & -To prevent contamination in culturing \tn 
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 & -Specific antibiotics - \seqsplit{penicillin/streptomycin} solutions \tn 
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\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Basic Constituents of Media (cont)}}  \tn
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 & -Broad spectrum antibiotics - kanamycin or amphotericin B \tn 
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 & -should not be toxic to the cells in culture \tn 
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 & -should depend on the type of contamination \tn 
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 & -Some antibiotics are used for selecting transfected animal cells, such as geneticin \tn 
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 & -\textgreater{}Transfected cell acquire resistant selectable marker and able to survive on medium containing geneticin \tn 
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 & -\textgreater{}Non-transfected cell could not survive \tn 
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10. Serum & -supernatant of clotted blood \tn 
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 & -Many undefined growth factors are contained, including many proteins and metal ions. \tn 
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\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Types of Serum}}  \tn
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\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650895536_Screenshot 2022-04-25 215011.png}}} \tn 
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\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Regulation of pH}}  \tn
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\mymulticolumn{1}{x{8.4cm}}{Regulation of pH is usually achieved by one of two buffering systems} \tn 
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\mymulticolumn{1}{x{8.4cm}}{1. "natural" buffering system} \tn 
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% Row 2
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\mymulticolumn{1}{x{8.4cm}}{-Gaseous CO2 balances with the CO3/HCO3 content of the culture medium} \tn 
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% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-Bicarbonate buffer requires 5-10 \% CO2 (supplied in a CO2 incubator) to maintain pH 6.9-7.4} \tn 
% Row Count 7 (+ 2)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-Bicarbonate/CO2 is low cost, non-toxic and also provides other chemical benefits to the cells.} \tn 
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% Row 5
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{2. chemical buffering using a zwitterion called HEPES \seqsplit{(4-(2-hydroxyethyl)-1-piperazineethanesulfonic} acid)} \tn 
% Row Count 12 (+ 3)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-HEPES has superior buffering capacity in the pH range 7.2-7.4} \tn 
% Row Count 14 (+ 2)
% Row 7
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-but is relatively expensive and can be toxic to some cell types at higher concentrations (above \textasciitilde{}100nMolar).} \tn 
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% Row 8
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\mymulticolumn{1}{x{8.4cm}}{-HEPES buffered cultures do not require a controlled gaseous atmosphere.} \tn 
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\begin{tabularx}{8.4cm}{X}
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Use of the phenol red indicator in media}}  \tn
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\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650895498_Screenshot 2022-04-25 214458.png}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
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\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Pros and Cons of Serum in Medium}}  \tn
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Advantage & Disadvantage \tn 
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1. Serum binds and neutralizes toxins & 1. It is not chemically defined, its composition varies a lot \tn 
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2. Serum contains a complete set of essential growth factors, hormones, attachment and spreading factors, binding and transport proteins & 2. It is sometimes source of contamination by viruses, mycoplasma, prions, etc. \tn 
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\SetRowColor{white}
3. Serum contains the protease inhibitors & 3. Sera could contain inhibitors or toxins \tn 
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4. Serum increases the buffering capacity & 4. Some ingredients could cause unwanted reactions \tn 
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\mymulticolumn{2}{x{8.4cm}}{5. Serum provides trace elements} \tn 
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\hhline{>{\arrayrulecolor{DarkBackground}}--}
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\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Serum-free media (SFM)}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-A defined SFM is one in which a group of components of known purity, present at a known concentration, are formulated together to optimize performance for a given cell type} \tn 
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% Row 1
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\mymulticolumn{1}{x{8.4cm}}{} \tn 
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% Row 2
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\mymulticolumn{1}{x{8.4cm}}{The two main approaches generally followed in designing a serum-free medium are:} \tn 
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\mymulticolumn{1}{x{8.4cm}}{1. Reduced serum} \tn 
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% Row 4
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\mymulticolumn{1}{x{8.4cm}}{-concentration of serum in the basal medium is progressively reduced whilst other components, e.g. growth factors and hormones, are added to identify the factors capable of restoring growth to the level obtained in the presence of serum} \tn 
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\mymulticolumn{1}{x{8.4cm}}{2. Basal medium} \tn 
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% Row 6
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\mymulticolumn{1}{x{8.4cm}}{-add components (singly or in combinations) to a basal medium in a stepwise manner until a medium is progressively 'built up, to give a similar or equivalent cell growth to the serum-supplemented medium} \tn 
% Row Count 18 (+ 5)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
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\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Pros and Cons of Serum Free Media}}  \tn
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Advantage & Disadvantage \tn 
% Row Count 1 (+ 1)
% Row 1
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1. Avoids qualitative and quantitative fluctuations of nutrients & 1. Multiplicity of media-Each cell type appears to require a different recipe \tn 
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2. Growth conditions are defined and controlled & 2. Selectivity-Some media may select a sublineage that is not typical of the whole population \tn 
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3. Small chances of contamination & 3. Reagent purity-The removal of serum also requires that the degree of purity of reagents and water and the degree of cleanliness of all apparatus be extremely high, as the removal of serum also removes the protective, detoxifying action that some serum proteins may have \tn 
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% Row 4
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4. More consistent performance & 4. Cell \seqsplit{proliferation-Growth} is often slower in serum-free media, and fewer generations are achieved with finite cell lines \tn 
% Row Count 31 (+ 7)
\end{tabularx}
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\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Pros and Cons of Serum Free Media (cont)}}  \tn
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\mymulticolumn{2}{x{8.4cm}}{5. Allows the possibility of studying the effects of one component present in serum, while eliminating the rest} \tn 
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\mymulticolumn{2}{x{8.4cm}}{6. Avoid ethical issues} \tn 
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\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
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% That's all folks
\end{multicols*}

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