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omgmbd > L5: Stem Cells & Regenerative Medicine > Flashcards

L5: Stem Cells & Regenerative Medicine Flashcards

1
Q

What are the types of stem cells?

A

→embryonic
→induced
→pluripotent

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2
Q

What is the origin of embryonic stem cells?

A

→pre-implantation embryo

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3
Q

Give examples of stem cells uses

A 
→ Model for basic and translational studies
→Cell replacement therapy
→Cell differentiation
→3D organoid models
→Drug screening
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4
Q

What can endodermal line differentiate into?

A

→lung

→pancreas

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5
Q

What can mesodermal line differentiate into?

A

→heart muscle

→RBCs

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6
Q

What can ectodermal line differentiate into?

A

→skin

→neuron

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7
Q

How are tissue specific stem cells maintained?

A

→special supportive microenvironments called stem cell niches.

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8
Q

What are the features of stem cell niches?

A

→Supporting ECM
→neighbouring niche cells
→secreted soluble signalling factors (e.g. growth factors and cytokines)

→physical parameters; shear stress, tissue stiffness, and topography),

→environmental signals (metabolites, hypoxia, inflammation, etc.).

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9
Q

What are the advantages of ESCs

A

→pluripotent- differentiate into any type of cell
→unlimited growth potential, high cell potency
→very low probability of mutation-induced damage in DNA

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10
Q

What are the disadvantages of ESCs?

A

→higher risk of tumour creation- spontaneous disease

→risk of being genetically different from the recipient’s cells- rejection

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11
Q

What are the advantages of adult stem cells?

A

→compatible with recipient’s cells- low risk of rejection

→less risk of tumour creation

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12
Q

What are the disadvantages of adult stem cells?

A

→oligopotent, limited cell potency
→limited numbers may be obtained
→higher probability of mutation induced damage of DNA

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13
Q

What are the advantages of induced pluripotent stem cells?

A

→low risk of rejection

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14
Q

What are the disadvantages of iPSCs?

A

→low growth potential
→low risk of tumour formation
→rather limited numbers may be induced
→higher probability of mutation induced damage of DNA

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15
Q

What do iPSCs and ESCs have in common?

A

→cells have a similar global gene expression profile to that of ES cells

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16
Q

Describe how iPSCs are generated?

A

→c-Myc promotes DNA replication and relaxes chromatin structure,

→allows Oct3/4 to access its target genes.

→Sox2 and Klf4 also co-operate with Oct3/4 to activate target genes
these encode transcription factors which establish the pluripotent transcription factor network

→result in the activation of the epigenetic processes (more open chromatin) that establish the pluripotent epigenome.

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17
Q

What are the pluripotent factors involved in iPSCs generation?

A

→c-Myc
→Oct3/4
→Sox2
→Klf4

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18
Q

What factor does Klf4 and Sox2co-operate with?

A

→Oct3/4

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19
Q

What type of chromatin is present in pluripotent iPSCs?

A

→most of the chromatin exists as euchromatin

→bearing histone marks associated with transcriptional activity

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20
Q

What is adult cardiomyocyte turnover like?

A

→low

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21
Q

What are the two types of regenerative strategies?

A

→Cell transplantation

→direct stimulation of endogenous cardiomyocyte production

22
Q

What is cell transplantation aimed at?

A

→replenishing lost cardiomyocytes.

23
Q

What are the challenges of cell based regenerative therapies?

A

→Immune rejection, manufacture

→isolation of sufficient cells, mode of delivery

→clinical regulation all challenges.

24
Q

What does cell free regenerative therapy involve?

A

→re-activation of developmental pathways e.g. epicardium based on models where the is no/reduced scarring and full cardiac regeneration

25
Q

What is neovascularisation?

A

→Improved circulation to injured area

→Paracrine effects improving CM replacement

26
Q

What is reactivation of epicardium important for?

A

→coronary blood vessels

27
Q

Compare immune response of CV disease in adults and neonatal

A

→adults= monocytes derived macrophage, no CM proliferation, and limited vascularisation
→fibrotic scar, contractile dysfunction

→neonatal= infiltration of injury by embryonic macrophages, and cardiomyocytes proliferation, and CM revascularisation
→functional recovery

28
Q

How are cardiac lineages from iPSCs made?

A

→Somatic cells are reprogrammed by Ymanak factors eg OCT4 and Sox2 to make iPSCs
→iPSCs exposed to GSK3b inhibition to make pre-cardiac mesoderm
→Wnt signalling inhibition to make CV progenitor cells
→exposed to TGF-b, PDGF, VEGF to make specialised cardiac cells

29
Q

What is necessary for epicardial migration and coronary vasculature in developmental gene activation?

A

→Myocardial thymosin β4

30
Q

What does addition of Tb4 to adult hearts do?

A

→stimulate epicardial outgrowth and neovascualarisation

31
Q

What is involved in transplantation and paracrine signals in CV regeneration?

A

→grafting sheet cells onto a heart
→cells do not seem to integrate into the heart tissue. Instead, they may release paracrine factors that help to regenerate the damaged muscle.

32
Q

What is lost in a MI?

A

→Epicardial expression

→FSTL1 in the epicardium has potent cardiogenic activity

33
Q

What is involved in stem cell therapy for cancer?

A

→Effector immune cells from iPSC/ESCs e.g. engineered T and NK cells targeted for immunotherapy.
→MSCs/NSCs deliver genes, nanoparticles, and oncolytic viruses to tumour niche
→Mutation correction in vitro, drug testing in vitro before replacement in vivo.

34
Q

How are burns treated?

A

→Generate ECM and produce paracrine signals which aid healing

35
Q

What are the different types of stem cells involved in burn therapy?

A

→fetal fibroblasts from ESCs
→epidermal stem cells
→mesenchymal
→iPSCs

36
Q

How are fetal fibroblasts used in burn therapy?

A

→improve skin repair due to the high expansion ability,
→low immunogenicity,
→intense secretion of bioactive substances such asFGFs, VEGFs, KGFs

37
Q

What do fetal fibroblasts secrete?

A

→FGFs, VEGFs, KGFs

38
Q

Why are epidermal stems cells used in burn therapy?

A

→high proliferation rate
→easy access
→keep their potency and differentiation potential for long periods.
→Generate most skin cell types for repair and regeneration

39
Q

Why are MSC used in burn therapy?

A

→They have a high differentiation potential
→a certain degree of plasticity.
→Migrate to the injured tissues, differentiate,
→regulate the tissue regeneration by the production of growth factors, cytokines, and chemokines

40
Q

Why are iPSCs used in burn therapy?

A

→can be differentiated into dermal fibroblasts, keratinocytes, and melanocytes

41
Q

What are limbal stem cells?

A

→Stem cells at the edge of the cornea

42
Q

What happens if limbal cells ate lost?

A

→cornea can no longer be repaired

43
Q

How are limbal cells used to treat eye injury or disease?

A

→Limbal stem cells are collected from an adequately healthy donor eye
→are expanded in the laboratory to sufficient number
→transplanted into the damaged eye
→Repairs the cornea and permanently restores vision.

44
Q

How is rejection avoided in eye injury therapy?

A

→works if the patient has a healthy section of limbus from which to collect the limbal stem cells.

45
Q

How is iPSCs used to treat eye injury?

A

be induced to make corneal epithelial cells for transplant

→exposure to the right signals can transform fibroblast cells into limbal stem cells

46
Q

What is RPE?

A

→Retinal pigment epithelium (RPE) is a single layer of post-mitotic cells, acting as a selective barrier to and a vegetative regulator of the overlying photoreceptor layer

47
Q

What can happen to the eye without RPE?

A

→parts of the retina can die

48
Q

How can the RPE be damaged?

A

→age-related macular degeneration (AMD),
→retinitis pigmentosa
→Leber’s congenital aneurosis.

49
Q

How can RPEs be made?

A

→ESC and iPSC

50
Q

How are iPSCs used to treat spinal injury?

A

→somatic cells biopsies from patients
→transform into iPSCs
→differentiate into different neural cells

51
Q

What results do NSPCs therapy in spinal injury show?

A

→can form functional synaptic subnetworks whose activity patterns resemble intact spinal cord
→grafts can integrate into sites of spinal cord injury (SCI) and generate neuronal relays across lesions

omgmbd (29 decks)

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