Embryonic stem cells Flashcards
What are XEN?
Extra embryonic stem cells
What are TS?
Trophoblast stem cells
What are mES and hES?
Mouse embryonic stem cells
Human embryonic stem cells
What are EpiSC?
- Epiblast stem cells
- Derived just before gastrulation (post-implantation)
Examples of pluripotent stem cells?
XEN TS mES hES EpiSC
What is the trophoblast?
Cells forming the outer layer of a blastocyst (the trophoectoderm)
Where have XEN and TS cells been derived from?
Mouse NOT the human
What are 8 properties of ES cells?
1) Derived from the inner cell mass of blastocysts
2) Non transfomed
- But high telomerase acvtivity
3) Indefinite proliferative potential
- High amplification potential
4) Stable diploid karyotype
5) Clonogenic
- Whole population can be re-derived from a single cell
6) Pluripotent
- Make all 3 germ layers
7) Carry particular markers
- Surface markers
- Genes
8) Can be incorporated into chimeras
- ONLY MOUSE ES CELLS (not human)
Which stem cells are naive and what does this mean?
Mouse ES cells
- Don’t express lineage markers
- Only express SC genes
Which stem cells are primed and what does this mean?
Mouse EpiSC and human PSC
- Express lineage markers
- Start to switch on other genes, as well as having SC genes turned on
How can you characterise PSC?
1) Surface markers
- Proteins on the surface
2) Genetically normal
- 46 XX or 46XY
3) Epigeneticically normal?
- Methylation
- Impriniting
4) Gene expression
5) Functional tests
- What can they do?
- Differentiation
- Clonogenic assays
How are antibodies used to characterise cells?
- Cells carry different combinations of marker proteins on their surface
- Different antibodies recognise specific proteins on the surface
- Probe them
What analysis techniques are involved in picturing antibodies?
Immunohistochemistry (homogenate)
Immunofluorescence (in situ)
What controls the dose of gene expression?
- Bilateral expression of genes
OR
- Maternal imprints or paternal imprints of genes
How are pluripotent stem cells different from eachother?
- Different characteristics depending upon where cultured from (mouse/human) and the timings
- Characteristics are: gene expression, surface markers, epigenetic and functional (differentiation potential and single-cell re-plating ability)