Please enable JavaScript.
Coggle requires JavaScript to display documents.
Key Concepts (Midterm) - Coggle Diagram
Key Concepts (Midterm)
Organoid Protocol (Week 6)
organoid protocol requirements
provision of a 3D environment conducive to cell aggregation & growth that allows for media exchange (nutrients in/ waste out) over a period of weeks while also being compatible with microscopy in some form
types of protocols
microwell culture
microchannel-based culture
liquid drop-assisted culture
centrifugation-based culture :
Disease Models (Week 5)
why use organoids to study disease?
disease may not be properly represented in cells
some diseased cells from humans may be impossible to study due to access / rarity
more drug combinations can be tested than in patients or mice
personalisation of organoids allows for testing of patient-specific approaches
types of disease models
infectious disease
hereditary disease
toxicology
cancer
History of Organoids (Week 2)
4 foundations of modern organoids
cells & self organisation
dissociated cells can self-organise back togeter
stem cells
increasing prominence of stem cells
new culture technologies
finding new ways to grow cells
extracellular matrix
discovering the extracellular matrix
Introduction to Cell Culture (Week 3)
cells for research
how are cells obtained for research?
primary cells
directly from patient
cell lines
immortalised primary cells that can proliferate indefinitely
popular cell types for research
cancer
breast cancer cells
HELA
lung cancer cells
immune
T cells
macrophages
fibroblasts
fibroblasts
cancer-associated fibroblasts
keratinocytes
mesenchynal stem cells
bone marrow-derived stem cells
adipose-derived stem cells
embryonic stem cells
human embryonic stem cells
induced pluripotent stem cells
how are cells most commonly grown?
adherent single cell (2D)
adherent monolayer (2D)
suspension
materials required for cell culture
containers
polysterene
flat surface
clear
cell culture media
nutrient provision
DMEM
FBS
energy provision
glucose
sodium pyruvate
antibiotics to prevent bacterial contamination
penicilin
pH indicator
phenol red
maintenance
sterile filtering
media changes
passaging
equipment
tissue culture hood / biosafety cabinet
cell incubator
centrifuge
water bath
limitations of standard cell culture
poor mimicry of in vivo environment
difficult to grow multiple cell types together
loss of 3D in vivo structural complexity
difficult to grow cells for a long time
Cell Sources & Tissue Goals (Week 4)
types of differentiation relevant to organoids
differentiation (mirroring development)
usage of progenitor cells (adult stem cells)
transdifferentiation
directly from one cell lineage to another
dedifferentiation
form progenitors (adult stem cells) or pluripotent cells (iPSCs)
methods to culture pluripotent stem cells
embryoid bodies
monolayer culture on ECM
culture on feeder layers
gastrulation
transition from single layer of epithelial cells (blastula) to multilayered 3D structure (gastrula)
begins with formation of primitive streak (PS)
consists of 3 layers: posterior, mid & anterior
all 3 layers functionally diff
brachyury is expressed everywhere
Foxa2 is only expressed at anterior end
gastrula
mesoderm
middle of 3 layers
forms heart, blood, vascular and skeletal muscle
ectoderm
outer layer of 3 germ layers
neural, skin
endoderm
innermost of 3 layers
eventually forms lung, liver, pancreas
types of organoids
pluripotent stem cells
activin a (nodal)
endoderm
intestine, stomach, liver
thyroid, lung
activin a (nodal) + bmp4
mesoderm
kidney
blood vessel
wnt, bmp4
ectoderm
brain organoids
adult stem cells
intestine, pancreas, lung, endometrium, stomach, prostate, liver