Cell Biology Mod. 1

Front 1

Commonality among diff. cells

Back 1

-genetic information

-similar structural features (e.g. membranes)

-carry out similar basic biochemistry and cell biology

Front 2

Similar basic biochem and cell bio

Back 2

-transmission of genetic infer

-basic metabolism: production & utilization of energy

-reproduction by cell division

-response to external and internal signals

Front 3

cellular diversity

Back 3

cells come in a variety of -shapes and sizes

-some are unicellular and some are multicellualr

Front 4

Cell Theory

Back 4

Applies to both prokaryotic cells and eukaryotic

-cell is the basic structural unit of life (contains hereditary info. and surrounded by semi-permeable membrane)

-cells arise only from preexisting cells; basic unit of reproduction

-organism is composed of one or more cells

Front 5

eubacteria

Back 5

relatively simple, have no nucleus

Front 6

archaea

Back 6

are prokaryotes, resemble prokaryote eubacteria in appearance and handling metabolism and energy conversion

resemble eukaryotes in genetic info transfer

Front 7

Diff. between prokaryotic cells and eukaryotic cells

Back 7

Eukaryotes have nucleus's and organelles (organelles are membrane-bound structures inside the cell)

Prokaryotes lack membrane bound organelles (has periplasmic space and systolic compartment)

Front 8

similarities between Gram + and Gram -

Back 8

-membrane proteins, inner membrane, periplasmic space (but larger in gram +

-Gram+ has one cytoplasm compartment, teichoic acid and thick peptidoglycan layer

-Gram - has two compartments, cytoplasm and periplasm, with a thin peptidoglycan layer in between, and LPS on outer membrane

Front 9

Cell culture general

Back 9

maintains cell shape

many of cell properties are maintained in culture both structural and functional

Front 10

tissue culture

Back 10

study the structural and functional properties of cells,

genetic and biiochemical manipulations

w/out interference from other cells

Front 11

what can you identify through tissue culture

Back 11

what nutrients, hormones, and growth factors in media are necessary to maintain growth rate, specialization, and cell death

what structural, function, and signaling changes occur

Front 12

genetic and biochemical manipulations

Back 12

easier to make antibodies and other important biomedical gene products

change cells for a therapeutic potential (reprogramming stem cells, tissue engineering)

change cells for a research potential

change cells for reproductive cloning

Front 13

proliferate

Back 13

grow

Front 14

differentiate

Back 14

cells specialize

Front 15

apoptosis

Back 15

cell death

removing old cells to stay young

Front 16

primary cells

Back 16

cells isolated from living organism

Front 17

tissue culture definition

Back 17

procedures used for maintenance and growth of cells or tissues in an artificial and aseptic environment

Front 18

cell culture definition

Back 18

isolated cells from tissues (organs) grown in culture

Front 19

organ culture (explant cultures)

Back 19

the culture of the whole or fragments of an organ

Front 20

use of primary cells

Back 20

can be immortalized by introducing telomerase gene, oncogene

can subculture (eventually will die by replicative cell senescence)

~very small amount of these cells survive as an immortalized
cell line

not always representative of original cell

Front 21

subculture

Back 21

use trypsin or EDTA to dissociate and detach cells from the plate, collect cells, and reseed cells onto a new plate

Front 22

Primary cell line characteristics

Back 22

have a finite lifespane: slows groth and undergo replicative cell senescence

"normal" chromosome number-maintained "normal" cell characteristics

have to isolate from living organism

Front 23

cell line characteristics

Back 23

immortalized cell line can grow continuously

cell line can have abnormal chromosome number-may not have normal cell characteristics

easier to access

Front 24

normal cell

Back 24

contact inhibition: cells slow the proliferation rate when they are touching and form a sheet of cells (monolayer)

anchorage-dependent: require glass or plastic surface and extracellular matrix proteins to grow

Front 25

Derivation of cell lines

Back 25

many cell lines are derived from tumors

all of them are capable of indefinite replication in culture and express at least some of the special ccharac. of cell's origin

Front 26

contact inhibition and confluence

Back 26

cells are confluent if rate of growth equals the rate of cell death and the total cell number is constant.

contact inhinition leads to a monolayer

growth of transformed cells are not regulated (cancer cells) continued to divide, forming foci of cells

Front 27

normal cell characteristics

Back 27

contact inhibited

reach confluency after a certain time

grow as a monolayer on a plastic dish

anchorage-dependent-grow on plastic coated w/ attachment factors

Front 28

transformed cell characteristics

Back 28

not contact inhibited

keep on growin and form foci

can grow as a multilayer on a plastic dish

anchorage independent, can grow in suspesion or in soft agar

need aeration

Front 29

advancement of cell culture

Back 29

the use of enzymes such as trypsin and collegenase to isolate cells from tissue

development of chemically defined, nutrient-rich media to grow cells

the use of antibiotics to control contamination

Front 30

tissue culture experiment

Back 30

removal of organ/tissue from animal/plants

cut and mince

dissociate the tissue with trypsin/collegenase enxymes to prepare isolated cell suspenstion

separate different cell types by size, shape, cell surface marker

plate/seed on plastic plates contianing medium (nutrient), plates could be coated w/ attachment facortor

put the plates into an incubator

take out plates and feed every other day or so

*depending on what you are starting with you may not have to do some of the steps

Front 31

tissue culture involves

Back 31

growing cells or tissue on medium(liquid or semi-solid) that supplies nutrient in an artificial, aseptic environment, which can be glass or plastic

cells are usually grown in an controlled environment, temp,

some cells require attachment factors

some cells do not require attachment factors and are grown as cell suspension in spinner bottles

Front 32

cell culture applications

Back 32

production of monoclonal antibodies and other specific products by cloning

Front 33

Cloning

Back 33

clone a specific gene into cultured cells to produce gene products

using cell fusion technology--> make hybridomas which are hybrid cells that produce monoclonal antibody

Front 34

Hybridoma

Back 34

know the stem in forming hybrid cells 1/11/12 pg 9 and 10

Front 35

formation of hybrid cells

Back 35

suspension of 2 cell types centrfuged w/ a fusing agent added

cell fusion and formation of heterocaryons, which are then cultured

selective medium allows only heterocaryons to survive and proliferate, these becom hybrid cells, which are then cloned

Front 36

antibodies

Back 36

can be used to detect proteins

2 binding sites (epitope recognition

*epitope=antigenic determinant

Front 37

polyclonal antibodies

Back 37

mixture of antibodies recognizing diff. antigenic determinants or epitopes

Front 38

monoclonal antibodies

Back 38

recognize one epitope on a protein

Front 39

3 different ways to make stem cells

Back 39

isolate pluripotent embryonic stem cells

make pluripotent ES cells by somatic nuclear transplantation

direct reprogrammin adult cells directly changed to induced pluripotent stem cells (iPS)

Front 40

pluripotent

Back 40

potential to differentiate into endoderm, mesoderm, or ectoderm

Front 41

ES cells from somatic cell nuclear transplantation

Back 41

fig 8-6

Front 42

3 components of biomembrane

Back 42

1. proteins
2. lipids
3. carbohydrates

Front 43

lipids and proteins

Back 43

are on all biomembranes (plasma and organelle)

Front 44

carbohydrates

Back 44

2-10% of biomembrane

covalently attached to lipids and proteins

on outers side of plasma membrane, also on luminal side of some endosomal membrane

Front 45

role of carbohydrates on glycolipids/glycoproteins

Back 45

-protect cells from physical and chemical stress

-increase solubility of proteins or lipids by adding a negative charge

-available to interact w/ peptide hormones, growth factors, extracellular matrix proteins, ab

Front 46

double membranes

Back 46

nucleus (endosomal membranes)

mitochondria

chloroplasts (has a 3rd-thylakoid membrane on the inside)

Front 47

single membranes

Back 47

endosomal membranes (share membranes by transport vesicles)
~rough ER
~Smooth ER
~Golgi stacks
~secretory vesicles
~lysosomes
~vacuoles in plants
peroxisomes
glyoxysomes in plants

Front 48

3 principle types of lipids

Back 48

1. phosophoglycerides
2. shingolipids
3. sterols

Front 49

phosphoglycerides

Back 49

phosphorylated alcohol or amino acid (ethanolamine, serene, choline, or inositol) is attached to hydroxyl group of glycerol

two fatty acid chains (even # carbons) are linked at carboy end to the hydroxyl gouts of glycerol by 2 ester bonds (one tail has double bond and other none)

decarboxylated PS is PE (PS has a net negative charge), PI is a signaling molecule

Front 50

cardiolipin

Back 50

type of phosphoglyceride

douple phospholipid (Diphosphotidylglycerol) on inner membrane of mitochondria

reduces membrane permeability to protons

Front 51

plasmalogen

Back 51

an ether phospholipid

abundant in *myelin*, heart, testis, and kidney tissue

Front 52

sphingolipids

Back 52

derivatives of sphingosine (an amino alcohol w/ a hydrocarbon chain) & fatty acyl chain connected to sphingosine by an amide bond

Front 53

sphingomyelin

Back 53

type of sphingolipid

has a phosphocholine as a head group

found in white layer of brain

Front 54

glycolipids cerebroside

Back 54

type of sphingolipid

has a glucose or galactose as a head group

Front 55

glycolipids ganglioside

Back 55

type of sphingolipid

has a complex sugars containing at least one sialic acid as a head group

Front 56

sterols

Back 56

cholesterol in animals, stigma sterol in plant, ergosterol in fungi

have 4 hydrocarbon rings (rigid and planar) & short hydrocarbon chain, both form hydrophobic tail

a single OH group at the C3 position is hydrophilic

cells that are unable to synthesize cholesterol usually lyse and die

none found in bacterial membranes

Front 57

brain lipids

Back 57

1. sphingolipids

2. cerebroside

3. ganglioside

Front 58

lipid composition

Back 58

fluidity of membrane depends on its *composition & temp*

plasma membrane of liver and rbc differ

composition for organelle membranes differ

lipid composition of PM membranes similar to each other than lipid composition from the organelle membranes

Front 59

lipid molecule

Back 59

amphipathic (hydrophilic polar group and a hydrophobic part)
~dual solubility

Membrane has two faces
1. Exoplasmic
2. cytosolic

Front 60

langmuir trough experiment

Back 60

working on simple oils, showed that they tend to spread over the surface of water. By measuring the original volume of oil and the final area it covered, he was able to calculate the thickness of the film.

can measure monolayer of membrane

showed lipid area is twice the outside area of rbc

[http://www.nature.com/scitable/topicpage/discovering-the-lipid-bilayer-14225438]

Front 61

fluid mosiac model

Back 61

membrane is lipid bilayer w/ lipids in a fluid state and prteins floating in the sea of lipids as discrete islands

lipids and proteins form a membrane spontaneously (non-covalent assembly)

Front 62

show non-covalent assembly of membrane

Back 62

do reconstitution experiment using mild detergents
~detergents disrupt lipid bilayer and it will reform to a liposome (bilayer) spontaneously

Front 63

above critical micelle concentration

Back 63

emergent mixed w/ water form micelle

Front 64

lower than critical micelle concentration

Back 64

detergent coats the membrane-spanning part of proteins/lipids, and solubilize proteins and lipids

Front 65

membrane fluidity

Back 65

length and number of double bonds in hydrocarbon chains affect thickness and fluidity

Front 66

disordered region of lipid bilayer

Back 66

thinner

more unsaturated tails don't pack well, more fluid like

shorter the tail the more fluid like

Front 67

ordered region of lipid bilayer

Back 67

more saturated, packed tightly and more gel like

thicker

longer the tail more gel like (higher Tm)

Front 68

Tm

Back 68

temp at which half membrane is in gel phase or fluid phase

Front 69

cholesterol

Back 69

important for membrane structure

dependent on temp
1. above Tm cholesterol stiffens and decreases permeability to more gel like

2. at Tm cholesterol pulls tails apart so membrane is more fluid like

has more influence on some lipids than others (lipid ordering effect on PC but not on SM)

makes PC less fluid like an membrane thicker

Front 70

lipid rafts

Back 70

distinctive lipid and protein composition then rest of membrane

rich in cholesterol, sphingolipids, phsphatidylchloine

has more saturated lipids (ordered region)

ticker membrane

rich in GPI-anchored proteins and proteins w/ longer transmembrane region

Front 71

4 types of lipid movements

Back 71

lateral diffusion

rotation

flexing

transverse diffusion

Front 72

lateral diffusion

Back 72

2 microns/1 second (rate)

Front 73

rotation

Back 73

hydrocarbon tails rotate

Front 74

flexing

Back 74

tails of inner side of membrane flex

Front 75

transverse diffusion

Back 75

flip-flop exchange of lipids between two leaflets

Front 76

scamblase/flippase

Back 76

enzymes required to flip-flop lipid molecules

only cholesterols can flip-flop without enzyme

Front 77

lipid rafts: cell function

Back 77

glycolipids and GPI-anchored proteins are on the ectoplasmic leaflet

important for membrane trafficking, as in forming caveolae with cave-in proteins and important for receptor-mediated cell signaling