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  /Title (ptc-c7-basic-animal-cell-culture.pdf)
  /Creator (Cheatography)
  /Author (woozing)
  /Subject (PTC - C7 (Basic Animal Cell Culture) Cheat Sheet)
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    \vspace{-2pt}\large{\bf{\textcolor{DarkBackground}{\textrm{PTC - C7 (Basic Animal Cell Culture) Cheat Sheet}}}} \\
    \normalsize{by \textcolor{DarkBackground}{woozing} via \textcolor{DarkBackground}{\uline{cheatography.com/146689/cs/31839/}}}
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  \mymulticolumn{1}{p{5.377cm}}{\bf\textcolor{white}{Cheat Sheet}}  \\
   \vspace{-2pt}Not Yet Published.\\
   Updated 25th April, 2022.\\
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Overview}}  \tn
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Application}}  \tn
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\mymulticolumn{1}{x{8.4cm}}{1. The study of basic cell biology, cell cycle mechanisms, specialized cell function, cell–cell and cell–matrix interactions.} \tn 
% Row Count 3 (+ 3)
% Row 1
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\mymulticolumn{1}{x{8.4cm}}{2. Toxicity testing to study the effects of new drugs.} \tn 
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% Row 2
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\mymulticolumn{1}{x{8.4cm}}{3. Gene therapy for replacing nonfunctional genes with functional gene-carrying cells.} \tn 
% Row Count 7 (+ 2)
% Row 3
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\mymulticolumn{1}{x{8.4cm}}{4. The characterization of cancer cells, the role of various chemicals, viruses, and radiation in cancer cells.} \tn 
% Row Count 10 (+ 3)
% Row 4
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\mymulticolumn{1}{x{8.4cm}}{5. Production of vaccines, mABs, and pharmaceutical drugs.} \tn 
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% Row 5
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{6. Production of viruses for use in vaccine production (e.g., chicken pox, polio, rabies, hepatitis B, and measles).} \tn 
% Row Count 15 (+ 3)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Explant Culture Procedure}}  \tn
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1. Obtaining the Explant & -obtained surgically using sterile equipment from mammals, rodents or avian organs or tissues \tn 
% Row Count 5 (+ 5)
% Row 1
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 & -ex 1: a piece of gingival tissue following tooth extraction can be removed as an explant to establish human gingival fibroblasts \tn 
% Row Count 12 (+ 7)
% Row 2
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 & -ex 2: a piece of adipose tissue can be used to establish mesenchymal stem cells \tn 
% Row Count 16 (+ 4)
% Row 3
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2. Cut and Clean the Explant & -place the explant in a petri dish containing around 1-2 mL of incomplete medium (medium without serum) \tn 
% Row Count 22 (+ 6)
% Row 4
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 & -using a sharp surgical blade, you can cut it (usually around 1×1 mm pieces) \tn 
% Row Count 26 (+ 4)
% Row 5
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 & -collect the pieces of explant using a sterile forceps and wash gently \tn 
% Row Count 30 (+ 4)
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Explant Culture Procedure (cont)}}  \tn
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 & -washing can be done by transferring pieces into a centrifuge tube containing around 0.5 mL of incomplete medium \tn 
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% Row 7
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 & -gently mix by pipetting the medium 4 to 5 times, and allow the pieces to settle down and remove the upper medium \tn 
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% Row 8
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 & -can be repeated 2 or 3 times \tn 
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% Row 9
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3. Culturing the Explants & -obtained explants are aseptically placed on a coated surface and allowed to attach to the surface in the presence of a rich culture medium \tn 
% Row Count 21 (+ 7)
% Row 10
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 & - medium ex: basal minimal media, Dulbecco's Modified Eagle Medium (DMEM) or Minimum Essential Medium Eagle (MEM) supplemented with 10-15\% serum \tn 
% Row Count 29 (+ 8)
% Row 11
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 & -cultured in standard tissue culture conditions (pH 7.2-7.4, temperature 37°C, 5\% CO2 and humidity) to allow for cell migration and proliferation \tn 
% Row Count 37 (+ 8)
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 & -change the media every 3 days without disturbing the explants \tn 
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% Row 13
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 & -depending upon the health and age of the tissue, cells emerge out of the explant within 15-30 days \tn 
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% Row 14
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 & -once outgrowth of cells starts from the explant, add 5 mL of medium to the flask in subsequent days \tn 
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% Row 15
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4. Once outgrowth of cells starts from the explant, add 5 mL of medium to the flask in subsequent days & -after the explants are completely surrounded by the cells, you can trypsinise the cells and subculture. \tn 
% Row Count 20 (+ 6)
% Row 16
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 & -it is better to use a lower concentration of trypsin (e.g. \textless{}0.25\% of trypsin for 5 min) \tn 
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% Row 17
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 & -choose an appropriate size of flask for seeding, depending on the total number of cells obtained \tn 
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Primary Culture}}  \tn
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\mymulticolumn{2}{x{8.4cm}}{-cultures prepared from tissues taken directly from animals} \tn 
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% Row 1
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1. Organ Culture & (google)-organ culture is able to accurately model functions of an organ in various states and conditions by the use of the actual in vitro organ itself \tn 
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 & -maintenance of a piece of tissue, a part of organ or a whole organ in vitro \tn 
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% Row 3
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2. Primary Cell Culture & -when taken tissue is dissociated, mechanically or enzymatically, into single cells which could be plated on a coated surface \tn 
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% Row 4
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3. Slice Tissue Culture & -referred to as explant or organotypic cultures \tn 
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% Row 5
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 & -small pieces of tissue of interest are simply allowed to attach to an appropriate substrate and are cultured in enriched media \tn 
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% Row 6
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4. Re-aggregate Culture & -dissociated cells is kept in suspension rather than allowed to settle on and attach to solid substrate \tn 
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 & -cells tend to re-aggregate into small balls \tn 
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 & -allowed cells cells to develop in three dimensions \tn 
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% Row 9
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5. Histotypic or histoculture & -culture of intact tissues \tn 
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\mymulticolumn{2}{x{8.4cm}}{(Google) - Histotypic culture is defined as three-dimensional culture of one cell type, while the term organotypic implies the interaction of two or more cell types from a complex tissue or organ.}  \tn 
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1. \seqsplit{Epithelial-Like} & -cells that are attached to a substrate and appear flattened and polygonal in shape \tn 
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% Row 1
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2. \seqsplit{Lymphoblast-Like} & -cells that do not attach normally to a substrate but remain in suspension with a spherical shape \tn 
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3. \seqsplit{Fibroblast-Like} & -cells that are attached to a substrate and appear elongated and bipolar, frequently forming swirls in heavy cultures \tn 
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% Row 3
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\mymulticolumn{2}{x{8.4cm}}{{\emph{It is important to remember that the culture conditions play an important role in determining shape and that many cell cultures are capable of exhibiting multiple morphologies.}}} \tn 
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1. Primary Cell Culture & -Adherent Cell Culture \tn 
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% Row 1
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 & -Suspension Cell Culture \tn 
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2. Secondary Cell Culture & - \tn 
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3. Cell Line & -Finite Cell Line \tn 
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 & -Continuous Cell Line \tn 
% Row Count 9 (+ 2)
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% Row 0
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\mymulticolumn{2}{x{8.4cm}}{-maintenance of growth of cells in culture medium using suitable glass or plastic containers} \tn 
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% Row 1
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\mymulticolumn{2}{x{8.4cm}}{-using the mechanical or enzymatic methods} \tn 
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\mymulticolumn{2}{x{8.4cm}}{-dissociated directly from the parental tissue (such as kidney, liver)} \tn 
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% Row 3
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\mymulticolumn{2}{x{8.4cm}}{-they will attach, divide and grow} \tn 
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\mymulticolumn{2}{x{8.4cm}}{} \tn 
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% Row 5
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\mymulticolumn{2}{x{8.4cm}}{{\bf{2 types of primary cell culture depending upon the kind of cells in culture}}} \tn 
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a) Anchorage Dependent /Adherent cells & -require attachment for cell growth \tn 
% Row Count 10 (+ 2)
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 & -monolayer culture system \tn 
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 & -usually derived from tissues of organs such as kidney where they are immobile and embedded in connective tissue \tn 
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 & (google)-have to be detached from surface before being subcultured \tn 
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 & (google)-growth limited to surface area \tn 
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% Row 11
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b) Suspension Culture/Anchorage Independent cells & -do not require attachment for cell growth/do not attach to the surface of the culture vessels \tn 
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 & -all suspension cultures are derived from cells of the blood system because these cells are also suspended in plasma in vitro e.g. lymphocytes \tn 
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\mymulticolumn{2}{x{8.4cm}}{-When a primary culture is sub-cultured, it becomes known as secondary culture or cell line.} \tn 
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\mymulticolumn{2}{x{8.4cm}}{-cell population derived from a primary culture at the first subculture} \tn 
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\mymulticolumn{2}{x{8.4cm}}{(google)-usually clonal, meaning that the entire population originated from a single common ancestor cell} \tn 
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\mymulticolumn{2}{x{8.4cm}}{-the term does not imply homogeneity or the degree to which a culture has been characterized} \tn 
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\mymulticolumn{2}{x{8.4cm}}{} \tn 
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\mymulticolumn{2}{x{8.4cm}}{{\emph{may be finite or continuous depending upon whether it has limited culture life span or it is immortal in culture}}} \tn 
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a) Finite Cell Lines & -cell lines which have a limited life span and go through a limited number of cell generations \tn 
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 & -growth rate is slow and doubling time is around 24-96 hours \tn 
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b) Continuous Cell Lines & -grow indefinitely \tn 
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% Row 8
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 & -cell lines transformed under laboratory conditions or in vitro culture conditions give rise to continuous cell lines \tn 
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% Row 9
\SetRowColor{white}
 & -growth rate is rapid and doubling time is 12-24 hours \tn 
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c) Transformed Cell Line & -cell lines obtained from tumor cells \tn 
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\SetRowColor{white}
d) Clonal Cell Line & -cells could be cloned in continuous cell lines to obtain genetically homogenous population \tn 
% Row Count 33 (+ 4)
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\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Pros and Cons of Finite Cell Lines}}  \tn
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\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Difference of Normal and Transformed Cells}}  \tn
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Normal Cells & Transformed Cells \tn 
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% Row 1
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1. Anchorage-dependent (except blood cells) & 1. \seqsplit{Nonanchorage-dependent} \tn 
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2. Density-dependent inhibition of proliferation & 2. No density-dependent inhibition of proliferation \tn 
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3. Mortal; Finite Cell Line & 3. Immortal; Continuous Cell Line \tn 
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\SetRowColor{LightBackground}
4. Contact Inhibition; Monolayer Culture & 4. No Contact Inhibition; Multilayer Culture \tn 
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\SetRowColor{white}
5. Dependent on external growth factor signals for proliferation & 5. May not need an external source of growth factors \tn 
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6. Greater retention of differentiated cellular function & 6. Typically loss of differentiated cellular function \tn 
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\SetRowColor{white}
 & -shorter population doubling time \tn 
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% Row 8
\SetRowColor{LightBackground}
 & -reduced substrate adhesion \tn 
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% Row 9
\SetRowColor{white}
 & -genetic instability (e.g. show heteroploidy and aneuploidy) \tn 
% Row Count 26 (+ 3)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Contact-Inhibition of Growth}}  \tn
\SetRowColor{LightBackground}
\mymulticolumn{1}{p{8.4cm}}{\vspace{1px}\centerline{\includegraphics[width=5.1cm]{/web/www.cheatography.com/public/uploads/woozing_1650863425_What-is-Contact-Inhibition.jpg}}} \tn 
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{x{2.8 cm} x{5.2 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Density-dependent Inhibition of Proliferation}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-reduction in proliferative activity that correlates with the attainment of confluency, that is，occupancy of all available attachment surface} \tn 
% Row Count 3 (+ 3)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-can occur before confluence is reached, and reflects diminished nutrient supply and the release of cell-derived factors (including waste products) into the medium} \tn 
% Row Count 7 (+ 4)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{} \tn 
% Row Count 7 (+ 0)
% Row 3
\SetRowColor{white}
{\bf{Saturation Density}} & -population density (cells/cm2) at the point when it reaches density-dependent inhibition of growth \tn 
% Row Count 11 (+ 4)
% Row 4
\SetRowColor{LightBackground}
 & -population density (cells/cm2) at the point when it reaches density-dependent inhibition of growth \tn 
% Row Count 15 (+ 4)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Cell Ageing in Culture}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-also known as In vitro cell senescence} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-involve progressive alterations in a number of cell characteristics} \tn 
% Row Count 3 (+ 2)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{} \tn 
% Row Count 3 (+ 0)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{Normal cell lines commonly have a finite lifespan, that is, they do not grow beyond a finite number of cell generations (population doublings).} \tn 
% Row Count 6 (+ 3)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-Eg, the lifespan of normal diploid fibroblasts is in the range of 50-70 population doubling.} \tn 
% Row Count 8 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Transformed Cells}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-cancerous cells} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-possess all six hallmarks of cancerous cells :} \tn 
% Row Count 2 (+ 1)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{1. Growth factor independency} \tn 
% Row Count 3 (+ 1)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{2. No response to growth inhibitors} \tn 
% Row Count 4 (+ 1)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{3. Evasion of apoptosis (Natural cell death)} \tn 
% Row Count 5 (+ 1)
% Row 5
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{4. Can promote angiogenesis (the development of new blood vessels)} \tn 
% Row Count 7 (+ 2)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{5. Unlimited proliferation - rapid increase} \tn 
% Row Count 8 (+ 1)
% Row 7
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{6. Invasive - tending to spread very quickly and undesirably or harmfully} \tn 
% Row Count 10 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Immortalization}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-Cell lines that have unlimited lifespan arc termed immortal or, preferably, continuous} \tn 
% Row Count 2 (+ 2)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{} \tn 
% Row Count 2 (+ 0)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{{\emph{the term immortalized and transformed are not synonymous}}} \tn 
% Row Count 4 (+ 2)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{Although infinite lifespan is generally considered to be a characteristic of transformed cells, not all continuous cell lines exhibit alterations in growth control attributed to cellular transformation.} \tn 
% Row Count 9 (+ 5)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{} \tn 
% Row Count 9 (+ 0)
% Row 5
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{{\bf{Immortalized Cells }}} \tn 
% Row Count 10 (+ 1)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-not yet cancerous, but have sufficient mutations to be able to be passaged forever, unlike non-transformed, non-immortalized cells, which all have a finite passage number} \tn 
% Row Count 14 (+ 4)
% Row 7
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-population of cells from a multicellular organism due to mutation, which can escape normal cellular senescence and keep undergoing division} \tn 
% Row Count 17 (+ 3)
% Row 8
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-this kind of cells can grow in vitro for prolonged periods} \tn 
% Row Count 19 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Cell Strain}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-describe a subcultured population selected on the basis of its expression of specific properties, functional characteristics, or markers} \tn 
% Row Count 3 (+ 3)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Clonal Culture / Clonal Selection}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-clone} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-establishment of a cultured cell population derived from a single cell} \tn 
% Row Count 3 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Sub-culturing (or passage)}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-Transfer or transplant cells of an ongoing culture to a new culture vessel so as to propagate the cell population or set up replicate cultures for study.} \tn 
% Row Count 4 (+ 4)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{-Subculturing or splitting cells is required to periodically provide fresh nutrients and growing space for continuously growing cell lines.} \tn 
% Row Count 7 (+ 3)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{-Such cultures may be called secondary cultures (first subculture from primary culture)} \tn 
% Row Count 9 (+ 2)
% Row 3
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{} \tn 
% Row Count 9 (+ 0)
% Row 4
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{Criteria for Subculturing} \tn 
% Row Count 10 (+ 1)
% Row 5
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{1. Cell concentration: should not exceed 1 x 10\textasciicircum{}6 cells/mL for most suspension-growing cells} \tn 
% Row Count 12 (+ 2)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{2. pH: which is linked to cell concentration, and declines as the cell concentration rises} \tn 
% Row Count 14 (+ 2)
% Row 7
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{3. Time since last subculture: should fit a regular schedule} \tn 
% Row Count 16 (+ 2)
% Row 8
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{4. Cell production requirements: for experimental or production purposes} \tn 
% Row Count 18 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Pros and Cons of Animal Cell Culture}}  \tn
% Row 0
\SetRowColor{LightBackground}
Advantage & Disadvantage \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
1. Controlled physiochemical environment (pH, temperature, osmotic pressure, O2, osmolarity etc.) & 1. Expertise is needed, so that behavior of cells in culture can be interpreted and regulated. \tn 
% Row Count 6 (+ 5)
% Row 2
\SetRowColor{LightBackground}
2. Controlled and defined physiological conditions - nutrient concentration, cell to cell interactions, hormonal control. & 2. Need of expertise and technical skill for the development, and regular use of tissue culture. \tn 
% Row Count 13 (+ 7)
% Row 3
\SetRowColor{white}
3. Homogeneity of cell types (achieved through serial passages)/ Homogenous genetic population & 3. Ten times more expensive for same quantity of animal tissue; therefore, reasons for its use should be compelling. \tn 
% Row Count 19 (+ 6)
% Row 4
\SetRowColor{LightBackground}
4. Economical, since smaller quantities of reagents are needed than in vivo. & 4. Unstable aneuploid chromosome constitution. \tn 
% Row Count 23 (+ 4)
% Row 5
\SetRowColor{white}
5. Legal, moral and ethical questions of animal experimentation are avoided. & 5. Cost factor is a major limitation. \tn 
% Row Count 27 (+ 4)
% Row 6
\SetRowColor{LightBackground}
6. Cost effective screening assays & -Establishment of infrastructure, equipment and other facilities are expensive. \tn 
% Row Count 31 (+ 4)
\end{tabularx}
\par\addvspace{1.3em}

\vfill
\columnbreak
\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Pros and Cons of Animal Cell Culture (cont)}}  \tn
% Row 7
\SetRowColor{LightBackground}
7. Easy production of biopharmaceuticals & -It is estimated that the cost of production of cells is about 10 times higher than direct use of animal tissues. \tn 
% Row Count 6 (+ 6)
% Row 8
\SetRowColor{white}
8. Available in adequate numbers to do chemical study & 6. Control of the environmental factors (pH, temperature, dissolved gases, disposal of biohazards) is not easy. \tn 
% Row Count 12 (+ 6)
% Row 9
\SetRowColor{LightBackground}
9. Easy to add genes (transfection) or regulate protein levels (RNAi) & 7. The native in vivo cells exist in a three- dimensional geometry while in in vitro tissue culture, the propagation of cells occurs on a two dimensional substrate. \tn 
% Row Count 21 (+ 9)
% Row 10
\SetRowColor{white}
 & -Due to this, the cell to cell interactive characters are lost. \tn 
% Row Count 25 (+ 4)
% Row 11
\SetRowColor{LightBackground}
 & 8. The cell lines may represent one or two types of cells from the native tissue while others may go unrepresented. \tn 
% Row Count 31 (+ 6)
\end{tabularx}
\par\addvspace{1.3em}

\vfill
\columnbreak
\begin{tabularx}{8.4cm}{x{4 cm} x{4 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Pros and Cons of Animal Cell Culture (cont)}}  \tn
% Row 12
\SetRowColor{LightBackground}
 & 9. Tissue culture techniques are associated with the differentiation i.e. loss of the characters of the tissue cells from which they were originally isolated. \tn 
% Row Count 8 (+ 8)
% Row 13
\SetRowColor{white}
 & -This happens due to adaptation and selection processes while culturing. \tn 
% Row Count 12 (+ 4)
% Row 14
\SetRowColor{LightBackground}
 & 10. Continuous cell lines may result in genetic instability of the cells. \tn 
% Row Count 16 (+ 4)
% Row 15
\SetRowColor{white}
 & -This may ultimately lead to heterogeneity of cells. \tn 
% Row Count 19 (+ 3)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{x{2.64 cm} x{5.36 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Growth Measuring Methods}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{1. Direct Methods} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
2. Cells & -Packed Cell Volume \tn 
% Row Count 2 (+ 1)
% Row 2
\SetRowColor{LightBackground}
 & -Cell count and viability \tn 
% Row Count 3 (+ 1)
% Row 3
\SetRowColor{white}
 & -Colony forming unit \tn 
% Row Count 4 (+ 1)
% Row 4
\SetRowColor{LightBackground}
 & -Optical density (OD) \tn 
% Row Count 5 (+ 1)
% Row 5
\SetRowColor{white}
3. Tissues & -Fresh weight and dry weight \tn 
% Row Count 7 (+ 2)
% Row 6
\SetRowColor{LightBackground}
4. Indirect Method & -Mostly used for large-scale cultures \tn 
% Row Count 9 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{x{5.04 cm} x{2.96 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Growth Observing}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{1. Increase in turbidity of cells} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
2. Increase in size of tissues/ explants & -swelling \tn 
% Row Count 3 (+ 2)
% Row 2
\SetRowColor{LightBackground}
 & -curling \tn 
% Row Count 4 (+ 1)
% Row 3
\SetRowColor{white}
 & \seqsplit{-proliferation} \tn 
% Row Count 5 (+ 1)
% Row 4
\SetRowColor{LightBackground}
3. Decrease in turbidity and size & -death \tn 
% Row Count 7 (+ 2)
% Row 5
\SetRowColor{white}
 & -apoptosis and necrosis \tn 
% Row Count 9 (+ 2)
% Row 6
\SetRowColor{LightBackground}
4. Microscopic observation & -Stereoscope \tn 
% Row Count 11 (+ 2)
% Row 7
\SetRowColor{white}
 & -Inverted microscope \tn 
% Row Count 13 (+ 2)
% Row 8
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{} \tn 
% Row Count 13 (+ 0)
% Row 9
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{{\emph{Necrosis is caused by factors external to the cell or tissue, such as infection. }}} \tn 
% Row Count 15 (+ 2)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{X}
\SetRowColor{DarkBackground}
\mymulticolumn{1}{x{8.4cm}}{\bf\textcolor{white}{Characterization of Cell Lines}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{1}{x{8.4cm}}{a) growth rate} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{1}{x{8.4cm}}{b) karyotyping (C11)} \tn 
% Row Count 2 (+ 1)
\hhline{>{\arrayrulecolor{DarkBackground}}-}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{x{2.32 cm} x{5.68 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Growth Curve}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-established taking into consideration the population doubling time, a lag time, and a saturation density of a particular cell line.} \tn 
% Row Count 3 (+ 3)
% Row 1
\SetRowColor{white}
1. Lag Phase & The time the cell population takes to recover from such sub culture, attach to the culture vessel and spread. \tn 
% Row Count 7 (+ 4)
% Row 2
\SetRowColor{LightBackground}
2. Log Phase & In this phase the cell number begins to increase exponentially. \tn 
% Row Count 10 (+ 3)
% Row 3
\SetRowColor{white}
3. Plateau Phase & During this phase, the growth rate slows or stops due to exhaustion of growth medium or confluency. \tn 
% Row Count 14 (+ 4)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}

\begin{tabularx}{8.4cm}{x{3.68 cm} x{4.32 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Bacterial Growth Curve}}  \tn
% Row 0
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-Unicellular organisms divide by binary fission} \tn 
% Row Count 1 (+ 1)
% Row 1
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-Each cell grows to full size, replicates its genetic material then divides into two identical daughter cells.} \tn 
% Row Count 4 (+ 3)
% Row 2
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{-By identical means, two cells divide into four, four into eight and so on, leading to an exponential increase in cell numbers: 1 → 2 → 4 → 8 →2\textasciicircum{}n} \tn 
% Row Count 8 (+ 4)
% Row 3
\SetRowColor{white}
\mymulticolumn{2}{x{8.4cm}}{-If we were to plot the number of cells in a population against time, we would get an exponential curve} \tn 
% Row Count 11 (+ 3)
% Row 4
\SetRowColor{LightBackground}
-Growth usually slows down due to: & a) supply of nutrients becoming exhausted \tn 
% Row Count 13 (+ 2)
% Row 5
\SetRowColor{white}
 & b) because metabolism leads to an accumulation of harmful waste substances \tn 
% Row Count 17 (+ 4)
% Row 6
\SetRowColor{LightBackground}
\mymulticolumn{2}{x{8.4cm}}{} \tn 
% Row Count 17 (+ 0)
% Row 7
\SetRowColor{white}
Lag Phase & -When an inoculum of bacteria is first introduced into some growth medium, it will probably require a period to adapt to its new surroundings \tn 
% Row Count 24 (+ 7)
% Row 8
\SetRowColor{LightBackground}
 & -When an inoculum of bacteria is first introduced into some growth medium, it will probably require a period to adapt to its new surroundings \tn 
% Row Count 31 (+ 7)
\end{tabularx}
\par\addvspace{1.3em}

\vfill
\columnbreak
\begin{tabularx}{8.4cm}{x{3.68 cm} x{4.32 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Bacterial Growth Curve (cont)}}  \tn
% Row 9
\SetRowColor{LightBackground}
 & -Eg, the carbon source in the medium is unfamiliar, the cells will need time to synthesise the necessary enzymes for its metabolism. \tn 
% Row Count 7 (+ 7)
% Row 10
\SetRowColor{white}
 & -Synthesize molecules needed for protein synthesis and enzymes required for cell division \tn 
% Row Count 12 (+ 5)
% Row 11
\SetRowColor{LightBackground}
 & -no net increase in bacterial numbers, however the cells are metabolically active. \tn 
% Row Count 16 (+ 4)
% Row 12
\SetRowColor{white}
Length of the lag phase depend on: & a) age and general health of the cells in the inoculum \tn 
% Row Count 19 (+ 3)
% Row 13
\SetRowColor{LightBackground}
 & b) conditions of bacteria before transfer into growth medium \tn 
% Row Count 22 (+ 3)
% Row 14
\SetRowColor{white}
 & c) content of the growth medium \tn 
% Row Count 24 (+ 2)
% Row 15
\SetRowColor{LightBackground}
Log (exponential) Phase & -When the bacteria have acclimatized to their new environment and synthesized the enzymes needed to utilize the available substrates, they are able to start regular division by binary fission. \tn 
% Row Count 34 (+ 10)
\end{tabularx}
\par\addvspace{1.3em}

\vfill
\columnbreak
\begin{tabularx}{8.4cm}{x{3.68 cm} x{4.32 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Bacterial Growth Curve (cont)}}  \tn
% Row 16
\SetRowColor{LightBackground}
 & -leads to the exponential increase in numbers \tn 
% Row Count 3 (+ 3)
% Row 17
\SetRowColor{white}
 & -under optimal conditions, the population of cells will double in a constant and predictable length of time, known as the generation (doubling) time. \tn 
% Row Count 11 (+ 8)
% Row 18
\SetRowColor{LightBackground}
 & -Cells are dividing at maximal rate \tn 
% Row Count 13 (+ 2)
% Row 19
\SetRowColor{white}
 & -Cells are most susceptible to the action of antibiotics and other deleterious agents \tn 
% Row Count 18 (+ 5)
% Row 20
\SetRowColor{LightBackground}
Stationary phase & -exponential phase is limited by environmental factors, and as the rate of growth slows down, the culture enters the next phase \tn 
% Row Count 25 (+ 7)
% Row 21
\SetRowColor{white}
 & -The levelling out of the growth curve does not mean that cell division has ceased completely, but rather that the increase due to newly formed cells is cancelled out by a similar number of cell deaths. \tn 
% Row Count 35 (+ 10)
\end{tabularx}
\par\addvspace{1.3em}

\vfill
\columnbreak
\begin{tabularx}{8.4cm}{x{3.68 cm} x{4.32 cm} }
\SetRowColor{DarkBackground}
\mymulticolumn{2}{x{8.4cm}}{\bf\textcolor{white}{Bacterial Growth Curve (cont)}}  \tn
% Row 22
\SetRowColor{LightBackground}
 & -Occurs when the number of viable cells stops increasing \tn 
% Row Count 3 (+ 3)
% Row 23
\SetRowColor{white}
 & -Due to nutrients being used up and/or toxic products accumulating from cell's metabolism \tn 
% Row Count 8 (+ 5)
% Row 24
\SetRowColor{LightBackground}
 & -as the death rate increases, the overall numbers fall and we enter the final phase of growth. \tn 
% Row Count 13 (+ 5)
% Row 25
\SetRowColor{white}
Death (or decline) phase & -As cells die off and the culture is unable to replace them, the total population of viable cells falls. \tn 
% Row Count 18 (+ 5)
% Row 26
\SetRowColor{LightBackground}
 & -Exponential decrease in the number of viable cells \tn 
% Row Count 21 (+ 3)
\hhline{>{\arrayrulecolor{DarkBackground}}--}
\end{tabularx}
\par\addvspace{1.3em}


% That's all folks
\end{multicols*}

\end{document}