Experimental models for cell biology

Two conditions for any animal experimental model:

1- It should be easy to work with the model

2- We should already known the complete genome sequence

1-The Escherichia coli (E.Coli) bacteria: The simplest prokaryotic model

​ - Grows easily under certain laboratory conditions

​
​ - divides every 20 minutes
​
​ - Genome: 4.6 million base pair – 4000 genes
​
​ - Nutrients mixture for optimal growing:
glucose, salts, amino acids, nucleic acid precursors and vitamins
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​ - Nutrients mixture for limited growing:
glucose, salts and a source of nitrogen

What we have learnt from E.Colí? - DNA replication

- The genetic code

- Gene expression

- Protein synthesis

- Amino acids and nucleic acid biosynthetic pathways

2- Yeasts, the simplest model for eukaryotic cells (Saccharomyces cerevisiae)

​ - More complex, but still grow easily under certain laboratory conditions
​ - divides every 2 hours
​ - Genome: 12 million base pair – 6000 genes
​ - It contains nuclear membrane and subcellular organelles
​ - The DNA is organized in 16 linear chromosomes

From this model we learnt how an eukaryotic cell works:

- DNA replication, RNA processing, protein sorting and the regulation of cell división.
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3- Caenorhabditis elegans (c.elegans) A multicellular nematode (a worm)

​ - It grows easily and can be genetically modified

​ - divides every 2 hours
​ - Genome: 100 million base pair – 19.000 genes
​ - Adult worms have 959 somatic cells
​ - It has been extensively studied under the microscope.
​ - It is very interesting model for animal development and cell
differentiation
​

4. Drosophila melanogaster

- Genome: 180 millions base pair - 14,000 genes - Reproduction cycle: two weeks

Very useful organism for genetic experiments, the relationship between genotype and phenotype
has been extensively studied in this organism.

5. The Zebrafish (26000 genes)

- They can be easily maintained in a laboratory

- They reproduce quite rapidly (every three to four months)

- The embryos develop outside of the mother and are transparent, so, first steps in development can
be easily observed

- Many mutations have been already identified.

A very interesting model to study the cardiac system .

6. The mouse

​ - Mice and men share about 97.5% of their genes (Around 30.000 codifying genes)
​ - Mice can be genetically modified (with technical difficulties)
​ - The generation of new mice with mutant genes

​ The mouse is a great model to study physiology

7. Animal Cell culture

- Isolated cells in culture can be manipulated under controlled conditions.

- Signaling mechanisms controlling cell functions

Two types

1- Primary cell cultures: Somatic terminal differentiated cells (muscle cells and neurons)

2- Cell lines: Embryonic stem cells or cells derived from tumors

Cell culture is the technique of growing cells in the laboratory outside an organism. It allows
for a steady supply of cells to experiment on. Cells can be grown under different
experimental conditions to mimic disease states in the body. The differences in intracellular
signaling can be observed to better understand the pathology and physiology of the body.

Type of cells
Fibroblast-like cells: bipolar or multipolar, elongated in shape and grow attached to the plate
(adherent)

Epithelial-like cells: polygonal, with regular and adherent dimensions forming groups

Lymphoblast-like cells: spherical forms that grow in suspension (non-adherent)

Cell culture media
Composition of culture media for mammalian cells (ex. Dulbecco's Modified Eagle's Medium -
DMEM)

Experimental conditions

Culture media:

Incubation:

​ Incubation temperature: 37oC

​ Incubation atmosphere: 5% CO2, to allow an exchange with the bicarbonate of the medium
and thus regulate a pH=7
​ Humidity: 95%