Culture Media for growing cells

Slides:



Advertisements
Similar presentations
Chapter 6.4: The Building Blocks of Life
Advertisements

Biochemistry.
Cell Nutrients Nutrients required by cells can be classified in two categories: - are needed in concentrations larger than M. C, N, O, H, S, P, Mg.
Section 6.3 – Life Substances
2.3 Carbon Compounds Standard B.1.1
What type of food? Carbohydrates, protein, lipids?
Biochemistry Chapter 3.
Chapter 3 Biochemistry.
Basic Vocabulary  Monomer – basic unit of a polymer  Polymer – Large molecule composed of repeating basic units or monomers.
The Building Blocks of Life
Organisms are made up of carbon-based molecules.
Organic Compounds (Biological Molecules) © Lisa Michalek.
1 Biology 12 Molecules of Life – organic biochemistry b b To be considered organic, molecules must contain Carbon and Hydrogen atoms. Practice; organic.
Biomolecules  Form = Function  Review inquiry activity from previous class.
Organic Chemistry Notes All organic compounds contain carbon. Carbon is able to form covalent bonds with other carbon atoms and many other elements easily.
Carbon Compounds Chapter 2 Section 3 Part 1. Objectives  Describe the unique qualities of carbon  Describe the structures and functions of each of the.
Carbon Compounds Chapter 2 sec. 3. carbon Organic compounds contain carbon.
1 Functional groups These groups are hydrophilic and typically polar with the exception of the phosphate group and therefore increase the solubility of.
Bio-Chemistry Biology 1. Carbon – is found in all living things. 4 electrons in its outer energy level Carbon has the ability to bond with many different.
CHAPTER 6 SECTION 4 Macro molecules. Organic Chemistry The element carbon (C) is a component of almost all biological molecules Life on earth is often.
Organic Chemistry The element carbon (C) is a component of almost
Macromolecules Biology. What does organic mean?  Organic Molecules contain both carbon and hydrogen.  Inorganic Molecules - the rest.  Water  Salt.
BIOMOLECULES MACROMOLECULES  made by polymerization- large compounds built by joining smaller ones together.  Smaller units (subunits) are called monomers.
Aim: How are organic compounds important to living things?
Macromolecules! United Streaming Intro Video. Chemistry of Carbon Carbon can bond with many elements, including hydrogen, oxygen, phosphorous, sulfur,
The Molecules of Cells Section 2.3. Macromolecules Carbohydrates Proteins Lipids Nucleic Acids All organic (contain carbon). Only produced in living things.
Biochemistry. Please pick up a copy of the notes and write down the homework assignment.
Macromolecules The Building Blocks of Life
Conditions for Cell Culture
Ch.2-3 Carbon Compounds and Macromolecules
The Building Blocks of Life
Organic Compounds Essential Standard
Ch.3-1 Cell Processes Chemistry of life.
Chapter 2 – Molecules.
ORGANIC CHEMISTRY: Intro to Macromolecules
6/16/2018 Outline 2-3 Carbon Compounds 6/16/2018.
Organic Chemistry: “Chemistry of Life”
Macromolecules.
Chemistry of Living Things
Macromolecules The Building Blocks of Life
Carbon is the Main Ingredient of Organic Molecules
Biochemistry The Chemistry of Living Things
Biochemistry This is a typical protein… ’s of amino acids covalently bonded together into a knot like structure we call a globular shape.
C-Notes: Chemistry of Living Things (Biochemistry)
Chapter 6 Section 4: The building Blocks of life Ms Su’ad
Organic Compounds Essential Standard
Biological Macromolecules
Organic Compounds Biochemistry
The Building Blocks of Life
Major Organic Compounds
Unit 2 Part 1: Organic Compounds (Biomolecules) and Enzymes
Chemistry and Biomolecules
Organic Chemistry: “Chemistry of Life”
Review Water is __________________
Ch.2-3 Carbon Compounds and Macromolecules
Organic Chemistry: “Chemistry of Life”
The Building Blocks of Life
Unit D: Humans Systems.
one glycerol and three fatty acids Fats and oils
Organic Chemistry Chapter 6.
Bellringer We will be going over your Atoms, Ions, Molecules and Properties of Water Quiz today as the warm-up.
Biochemistry Lysozyme – a protein.
2.3: Macromolecules.
Chapter 2 Chemical Basis of Life
Vocabulary-Concept Review
Carbon, Important to life… Carbon is always moving through all living things, and is in every organic compound on earth. Just like water, nitrogen,
Macromolecules aka Giant Molecules.
Biological Compounds You are what you eat.
Biochemistry.
How does structure relation to function?
Presentation transcript:

Culture Media for growing cells

Animal cell. Animal cells do not have a cell wall and, thus, do not have a rigid cell boundary. The shapes of animal cells are quite diverse due to the flexibility of the outer membrane and the response when cells touch each other. Plant cell. Most plant cells contain chloroplasts and a rigid cell wall. Animal cells do not.

Each rod-shaped structure in this electron micrograph is an E Each rod-shaped structure in this electron micrograph is an E. coli cell. E. coli cells are simple prokaryotes with no membrane-bound organelles, such as mitochondria or chloroplasts. Shown in cell culture, these CHO cells are a common mammalian cell line used to manufacture recombinant protein.

The Molecules of Cells Engineered molecules are the basis of many biotechnology products. Cells are composed of a variety of molecules. Many molecules found in cells are much larger than atoms. Very large molecules are found in structural components.

Carbohydrates Carbon, hydrogen, and oxygen Ratio 1:2:1 Polysaccharides Excellent structural and energy-storing molecules Plants store glucose in starch molecules Monosaccharides Monomer units that cells use to build polysaccharides Most well known is glucose; an energy molecule

Disaccharides Produced when enzymes form a bond between two monosacchrides Sucrose is made when fructose and glucose are chemically combined Lipids Often referred to as hydrocarbons Three groups of lipids Triglycerides Phospholipids Steroids

Proteins Nucleic Acids The most important of the cellular molecules Nine different categories of proteins Structural Enzyme Transport Contractile Hormone Antibody Pigment Recognition Toxins A typical cell produces more than 2000 proteins Amino acids are the monomers of proteins There are 20 different amino acids found in proteins The fourth major group of macromolecules Two types of nucleic acids DNA RNA

Bacterial Culture Media Liquid vs. Solid/Semisolid Chemically defined vs. Complex Selective and Differential Media

Nutrient Requirements for Bacteria Element % of Dry weight Function Media source Carbon 50 Main building block Sugars, extracts Oxygen 20 Electron acceptor, water Air Nitrogen 14 Amino acids, nucleic acids, coenzymes Amino acids, extracts Hydrogen 8 Organic compounds and water Present in most chemicals added to media Phosphorus 3 Nucleic acids Na and K phosphates Sulfur 1 Amino acids & coenzymes Na sulfate Minerals (Na, K, Mg, Ca, Fe etc.) 2 – 4 Transport, cofactors etc. Salts Trace Elements Trace Variety of functions Salts of Co, Zn, Mo, Cu, Ni, Se, etc. Organic growth factors Growth factor Vitamins, purines, pyrimidines

Sources of Nutrients Yeast Extract Peptones Glucose (or other defined carbon sources such as acetate, glycerol, lipids etc.) Ammonium salts (for nitrogen) Mineral salts for trace elements (e.g. CaCl2, CuSO4 etc.) Growth factors ( e.g. vitamins, purines, pyrimidines etc.) needed for more fastidious bacteria Other supplements: antibiotics, pH indicators etc.

Method of making bacterial culture media Mix all ingredients in purified water Adjust pH if necessary Sterilize by autoclaving at 121oC for 15 minutes Cool medium to 55oC before adding sterile stock of antibiotic before pouring agar plates Some components may need to be autoclaved separately to avoid precipitation or unwanted chemical reactions

Nutrient Requirements for Cultured Mammalian Cells Component Function Media source Carbohydrates & derivatives Primary energy source Glucose Bulk ions and trace elements Transport, adhesion, signaling, cofactors etc. Salts of Ca, Cu, K, Na, Fe, Mg, Zn etc. Amino Acids Nitrogen source Amino acids Vitamins & coenzymes Enzymatic cofactors Vitamins (e.g. Biotin, folic acid, Vitamin B12 etc.) Lipids and derivatives Components of cell membrane Lipoic and linoleic acids, inositol, choline chloride Purines & pyrimidines Nucleic acid precursurs Hypoxanthine, thymidineHCl Proteins Growth factors, cytokines, attachment & transport Fetal Bovine Serum (FBS) or several proteins Non-hormonal & Non-nutritional supplements Buffers, antibiotics, pH indicators etc. Bicarbonate, pen/strep, phenol red

Media Selection: Commonly available basal media Media Type Common Uses Minimum Essential Medium (MEM) Broad spectrum Basal Media Eagle (BME) Diploid/ primary cells Dulbecco’s Modified Eagle (DME) CMRL (?) Earl’s “L”, monkey kedney cells Iscove’s Modified Dulbecco’s (IMDM) Rapidly proliferating high density cultures McCoy’s Human lymphocytes Ham’s F10 and F12 CHO cells RPMI 1630/1640 Suspension cells 11/11/2018

Making Media for Cell Culture Basal media is store bought Liquid, powder, or concentrate Add Serum, antibiotics, additional amino acids etc. Filter sterilize with 0.2 or 0.1 micron filter into pre-sterilized bottles