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Lecture 1 - Introduction to Cell Biology.docx

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Western University
Biology 2382B
Robert Cumming

Intro to Cell Biology – Jan 9 2013 Why Study Cells?  Perspective of cells simply as storehouse for thousands of genes  Function of cells helps to establish what is “normal” so as to be able to fix what is abnormal What is a Cell?  Ultimate goal – to understand how macromolecular systems and organelles work and cooperate to enable cells to function autonomously and in tissues How do we study cells?  Hypothesis-Driven experiments o First you need the tools and methods to ISOLATE and MAINTAIN cells in vitro To know how to view cells and what to look for o How to identify and study how proteins drive a cells biological process Cell Culture  Cell culture is the technique used to grow cells or tissues outside the organism under strictly controlled conditions  Cells are isolated from any tissue by BREAKING DOWN the cell-cell and cell- matrix interactions (Remember that cells like to stick together)  Can use mechanical fragmentation – using filter  trypsin (A protease which acts like a little lawn mower which will go around the cell and chew apart the extracellular proteins which make the outside of the cells so sticky  EDTA – Gets rid of the stickiness of the cell via extracellular interaction  Cells are supplied with proper nutrients o Amino acids, minerals, vitamins, salts, glucose, etc.  Serum o Insulin, growth factors  Grown at usually 37c in CO incubator 2  Cells can grow as adherent cell cultures or suspension cell cultures o Growth factors stimulate cells to divide  Red in the tubes is phenol red which tells you the pH of the medium (If it’s yellow than you have acidic pH)  Generally grow cultures in petri dishes or culture flasks  The bottom of the culture flasks has a charge or protein which will allow the cells to stick (sometimes of course though you don’t want them to stick  Suspensions cell cultures o Spinner flasks – This causes mechanical agitation o Roller flasks – Constantly rolling o Both achieve cel cultures which don’t allow the cells to settle at the bottom (This would be ideal in the case of stem cells for example Intro to Cell Biology – Jan 9 2013 Cell Culture Cont.  Confluent monolayer – Contact Inhibition o Means that you have a single layer of cells across a surface – no longer piling on top of each other o Confluent means that they are NOT growing anymore – The cells sense that they are really close to each other and will stop growing o You can treat these cells with trypsin – this will cuase the cells to fall of the bottom of the dish and you can dilute and spread them out into different dishes and repeat the process of regrowing them to confluent monolayer stage o You can only do this a finite number of times though – only about 50 times  Primary Cell Culture o Refers to cells taken directly from an organism . These cells usually divide a limited number of times (~50 generations Hayflic limit). Also undergo contact inhibition if cell desnity is high  Cell Line o Refers to cells, which are transformed and are able to grow indefinitely. Also known as immortal cells or transformed cells (They’ve lost some of the feedback mechanisms to tell them to stop). Less likely to exhibit contact inhibition – these will grown ON TOP of each other  The first human cell line called HeLa was established in 1952 by George Gey – From cervical carcinoma biopsy of Henrietta lacks Morphology of Normal vs Transformed Fibroblast  You can try and take normal fibroblast and transform them by implementing new genes (it will disrupt normal cell cycling)  Normal o Elongated o Aligned o Orderly Paced o Grow in Parallel arrays o Contact Inhibition  Rounded o Not ordered o No contact inhibition  This is all to say that you can take normal cells and turn it into cell line Birth, Lineage and Death of Cells  All Cells start as STEM CELLS o Stem cells have unique properties – Can divde into two identical cells (Symmetric divison) but both of these can have a completely different fate Intro to Cell Biology – Jan 9 2013  As the cells begin to differentiate they go through assymettric division and can split into two cells of different function – These are generally later stem cells  These final differentiated cells are mature cells  Stem cells
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