MPharm4 SL52117 > Future Pharma > Flashcards

Future Pharma Flashcards

(45 cards)

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1
Q

What are stem cells

A

unspecialised cells with the ability to self-renew, producing more stem cells and differentiating into different types

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2
Q

What are the 3 different types of stem cells?

A

Embryonic
Induced pluripotent
Adult (somatic/tissue)

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3
Q

What are the 4 potencies of stem cells?

A

Totipotent
Pluripotent
Multipotent
Unipotent

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4
Q

What are the two main methods of using stem cells?

A
  • infusing differentiated cells into the body
  • form mini organs and allow to grow
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5
Q

Stem cell markers

A

Stem cells are detected by expression or absence of transcription factors
Markers can be used to enrich stem cell populations for further study or aplications like tissue engineering

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6
Q

What is used to determine expression?

A
  • western blotting
  • RT-PCR
  • immunocytochemistry
  • flow cytometry
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7
Q

What is used to enrich cell populations?

A
  • centrifugation
  • MACS
  • flow cytometry/FACS
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8
Q

What is density centrifugation?

A
  • centrifuge blood cells over a dense, carb-rich solution
  • density gradient separates cells, plasma, mononuclear cells (stem cells treated with specialised antibodies) then RBCs + granulocytes
  • magnetic-activated cell sorting (MACS) using magnetic nanoparticles
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9
Q

What is flow cytometry?

A
  • method for detecting specific molecules on + within cells
  • can also be used for sorting/isolating specific cells for analytical research
  • forward scatter vs side scatter
  • mainly used for proteins but can do RNA and DNA too
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10
Q

What is fluorescent activated cell sorting (FACS)?

A
  • adding antibodies on different antigens to separate cells based on this, applies a charge and gets divided
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11
Q

Embryonic stem cells

A

Characterised by self-renewal, indefinite proliferation and pluripotency
- pluripotency regulated by complex set of interactions between extra cellular signalling pathways, transcription factoors, mRNAs, epigenetic factirs
- ethical issues, teratoma formation, allogenic

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12
Q

Induced pluripotent cells

A

unlimited proliferation, surface markers and gene expression alike to ESCs
- differentiate into cells of all trhee germ layers in vitro + teratomas
- no ethical issues when derived from patient
- directed differentiation

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13
Q

Parkinsons disease and stem cells

A

plurippotent stem cells can be differentiatde into midbrain dopaminergic neurons ro replace those lost in PD
- mDA neurons arise from the floor plate in the developing brain
- directed differentiations of PSCs can be achieved by replicating the signalling/contact conditions
- cells differentiate via a variety of progenitor cells

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14
Q

Cancer and stem cells

A

induced pluripotent stem cells express a number of tumour-associatd antigens
- when tested against murine tumour models, vaccination with iPSCs + CpG led to tumour regression
- autologous iPSCs could act as personalised cancer vaccines - treatment possible within weeks of diagnosis
- advantages over current immunotherapy strategies

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15
Q

Transplant rejection

A
  • The human leukocyte antigen complex dtermines wherther our immune system sees cells as slef or non-slef
  • HLAs are co-dominantely expressed, inherited as haplotypes
  • if therapeutic cells arent HLA matched, they can be rejected (autologous cells ok)
  • immunosuppression necessary to prevent rejection
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16
Q

Immune compatibility of hPSCs

A
  • hPSCs were intially thought to be immune privegleged due to undifferentated state
  • more recent studies show this isn’t the casd
  • hPSCs express HLA class I antigens but not class 2
  • AB(O)H blood group antigen correlates to ABO genotype
    carefult HLA matching required
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17
Q

Hypoimmunogenic iPSCs

A
  • studying the interface between mother and foetus revealed molecules which may be involved in immune shielding
  • ## maintained iPSCs characteristics
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18
Q

Autologous

A

From the same individual

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19
Q

Allogenic

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A

From a donor source

20
Q

Haematopoiesis

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A

The production of blood cells
- occurs under steady state conditions,, in a healthy adult up to 5x10^11 cells a day
- all adults blood cells are descended from rare population of cells, HSCs which are able to self renew and give rise to diffferentiated cells of all haematopoietic lineages
- predominantly occurs in the red bone marrow

21
Q

Haematopoietic stem cells

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A
  • HSCs are the most studied and best characterised of all adult stem cells
  • adult humans believed to have around 50k to 200k active HSCs
  • they divide every 40 weeks
22
Q

Transplantation

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A
  • currently the only extablished stem cells therapy
  • autologous or allogenic depending on indication
23
Q

Process of transplanting autologous stem cells

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A
  • patient treated with drugs to increase numnber of stem cells
  • stem cells are collected from blood or marrow
  • blood is processed though a machine, cell removed and blood returned
  • patient recieves high dose chemo, to kill any remaining cancer cells
  • stem cells reinfused into patient and begin producing new blood cells
24
Q

Sickle cell and stem cells

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A
  • a single nucleotide polymorphism (A>T) in beta-globin gene
  • autologous stem cells transfected with anti sickling beta-globin gene
  • repopulates patients haematopoietic space with cells making corrected blood cells
25
Mesenchymal stem cells
- from bone marrow, aspirate with bone needle and centrifugr - seprate peripheral blood mononuclear cells of the mononuclear fraction, seen in culture as spindly cells - readily undergo osteogenesis - aren't functionally equivalent, differences in surface markers, proliferation and differentiation and affected by age too
26
Main sources of MSCs
- bone marrow (sparse, 100% isolation efficiency, high differentiation potential) - adipose tissue (plentiful, 100% isolation efficacy, high differentiation potential - embilical cord blood (very sparse, 30% isolation efficacy, medium differentiation potential)
27
The rols of mesenchymal stem cells
- implicated in tissue homeostasis and repair - mechanisms are not fully understood, mostly in vitro - roles (daughter cells that differentiate for repair, homing to sites of injury, secretion of factors tht support wound repair by recrioting other cell types and modulating the immune response) - response mediated by complex interplay of soluable fators, ECM molecules + cell surface receptors - paracrine functions
28
Therapeautic application of mesenchymal stem cells
- despite lack of knowledge regarding in vivo biology of MSCs have been clinically used for a number of years - once regarded as hypo immungenic - have immunomodulatory functions (useful in treatemtn or prevention of GvHD) - potential fror delivery of therapeutic proteins
29
Issues with mesenchymal stem cells
- low number in bone marrow - slow growing - can spontaneously differentiate
30
MSCs after myocardial infarction
- transfuse cells after MI - home to area of damage - secretes paracrine factors that can attract cardiac progenitor cells + cardiac stem cells - also potential to transdifferentiate into endothelial cells and into cardiomyocytes to repair damage to cardiac tissue
31
Potential cancer treatments with MSCs
- effectively hypoimmunogenic, can home to injury (tumours), can promote growth or engineered to deliver killer payload - can deliver proteins in iduce apoptosis - can secrete factors to starve tumor cells by reducing angiogenesis - can be used to deliver mRNA used to down regulate growth of cancer cells - trojan horse with typical chemo
32
Cell based muscular distrophy treatment
muscle stem cells repair damage - in musclular dystrophy, actin filaments join the outside of the cell via dystrophin - mutations in dystrophin can stop this occurring so muscle injuryies can be repaired - constant stae of injury, so muscle stem cells constantly activated, eventually become senescent and no longer able to repair damage - x linked, 16x more likely in men - reduced muscle fibre integrity > increased CA2+ influx > activation of Ca2+ sensitive proteases > necrosis of msucle tissue
33
xenotransplantation
- growing organs in pigs (cornea, lung, kidneys, heart, liver, pancreas)
34
Basic concepts of tissue engineering
- cell isolation and enrichment/purification - expansion of cell number - seeding on a suitable scaffold maturation of tissue - implantaion in pateint
35
Tissue engineering scaffolds
- acts as 3d support for cell growth - ideally: biocompatible, biodegradable, porous, cytocompatible, me hanically apporpriate,
36
Types of polypeptide scaffolds
collagen gelatin fibronectin fibrin laminin
37
Types of polysaccharide scaffolds
- hyalouronic acid, alginate, chitosan
38
Types of bioceramics/bioactive scaffolds
- hydroxyapatite - bioactive glass - alumina - TiO2 - calcium phosphates
39
MEthods for scaffold formation
- compression - solvent casting particle leeching - freeze drying - spinning - electrospinning - 3d printing
40
Bioreactors
Specialized devices that create a controlled in vitro environment to cultivate cells, facilitating the development of functional, three-dimensional tissues for regenerative medicine - depending on the tissue, may apply some force (commpression for bine tissue, tensions for muscle tissue)
41
The body as a bioreactor
Calcium alginate hyrdogel injected into bone to create 'in vivo bioreactor' - non adhesive scaffold made from seaweed, poor for cell growth but stimulates stem cells to proliferate to fill the scaffold
42
Challenges of tissue engineering - vascularisiation
cells need to be within ~200um of blood vessel to survive to get around this - seed scaffold with endothelial cells - incorporate VEGF into scaffold - build scaffold around vascular bed ex vivo
43
3D bioprinting
- attempts to address 3D organisation of cells - challenges: cell density, resolution, vascularisation, innervation, mechanical integrity
44
How does bacterial resistance occur?
- many bacteria both beneficial and pathogenic only few are resistant - abx kills most, resistant will survive - resistant org has no competition so survives and grows - resistance can transfer to other organisms
45
Bacteriophage therapy
the use of specialized viruses to treat bacterial infections by killing specific pathogenic bacteria advantages - self-replicating where target bacteria are present - slef-evolving to overcome resistance - naroow-spectrum of activity - potential for personalised therapy disadvantages - immunogenicity - naroow spectrum - pk - regulatory standards + manufacture challenges
MPharm4 SL52117
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