Generation and characterisation of four multiple sclerosis iPSC lines from a single family

Fortune, Alastair J; Taylor, Bruce; Charlesworth, Jac; Burdon, Kathryn P.; Blackburn, Nicholas B.; Fletcher, Jessica L.; Mehta, Ashish; Young, Karen

doi:10.1016/j.scr.2022.102828

Cited by 4 publications

(3 citation statements)

References 3 publications

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“…Additional healthy control iPSC lines were used as specified in text: MNZTASi019-A (from a 53 year-old female donor); MNZTASi021-A (76 year-old male donor), and MNZTASi022-A (56 year-old female donor) were purchased from the MS Stem biobank (Menzies Institute for Medical Research, Hobart, Tasmania, Australia). MS Stem iPSCs were generated and characterised as previously described [ 19 , 20 ] with approval from the University of Tasmania Human Research Ethics Committee (Project H16915). All iPSC lines were shown to have karyotypically normal karyograms within 10 passages of use for experiments and were used between passage 5–35.…”

Section: Methodsmentioning

confidence: 99%

Induced pluripotent stem cell derived pericytes respond to mediators of proliferation and contractility

King,

Courtney,

Brown

et al. 2024

Stem Cell Res Ther

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Background Pericytes are multifunctional contractile cells that reside on capillaries. Pericytes are critical regulators of cerebral blood flow and blood–brain barrier function, and pericyte dysfunction may contribute to the pathophysiology of human neurological diseases including Alzheimers disease, multiple sclerosis, and stroke. Induced pluripotent stem cell (iPSC)-derived pericytes (iPericytes) are a promising tool for vascular research. However, it is unclear how iPericytes functionally compare to primary human brain vascular pericytes (HBVPs). Methods We differentiated iPSCs into iPericytes of either the mesoderm or neural crest lineage using established protocols. We compared iPericyte and HBVP morphologies, quantified gene expression by qPCR and bulk RNA sequencing, and visualised pericyte protein markers by immunocytochemistry. To determine whether the gene expression of neural crest iPericytes, mesoderm iPericytes or HBVPs correlated with their functional characteristics in vitro, we quantified EdU incorporation following exposure to the key pericyte mitogen, platelet derived growth factor (PDGF)-BB and, contraction and relaxation in response to the vasoconstrictor endothelin-1 or vasodilator adenosine, respectively. Results iPericytes were morphologically similar to HBVPs and expressed canonical pericyte markers. However, iPericytes had 1864 differentially expressed genes compared to HBVPs, while there were 797 genes differentially expressed between neural crest and mesoderm iPericytes. Consistent with the ability of HBVPs to respond to PDGF-BB signalling, PDGF-BB enhanced and a PDGF receptor-beta inhibitor impaired iPericyte proliferation. Administration of endothelin-1 led to iPericyte contraction and adenosine led to iPericyte relaxation, of a magnitude similar to the response evoked in HBVPs. We determined that neural crest iPericytes were less susceptible to PDGFR beta inhibition, but responded most robustly to vasoconstrictive mediators. Conclusions iPericytes express pericyte-associated genes and proteins and, exhibit an appropriate physiological response upon exposure to a key endogenous mitogen or vasoactive mediators. Therefore, the generation of functional iPericytes would be suitable for use in future investigations exploring pericyte function or dysfunction in neurological diseases.

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Section: Methodsmentioning

confidence: 99%

Induced pluripotent stem cell derived pericytes respond to mediators of proliferation and contractility

King,

Courtney,

Brown

et al. 2024

Stem Cell Res Ther

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show abstract

“…Additional healthy control iPSC lines were used as specified in text: MNZTASi019-A (from a 53 year-old female donor); MNZTASi021-A (76 year-old male donor), and MNZTASi022-A (56 year-old female donor) were purchased from the MS Stem biobank (Menzies Institute for Medical Research, Hobart, Tasmania, Australia). MS Stem iPSCs were generated and characterised as previously described (18, 19) with approval from the University of Tasmania Human Research Ethics Committee (Project H16915). All iPSC lines were shown to have karyotypically normal karyograms within 10 passages of use for experiments and were used between passage 5-35.…”

Section: Methodsmentioning

confidence: 99%

Induced pluripotent stem cell derived pericytes respond to endogenous mediators of proliferation and contractility

King,

Courtney,

Brown

et al. 2023

Preprint

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BackgroundPericytes are multifunctional contractile cells that reside on capillaries. Pericytes are critical regulators of cerebral blood flow and blood-brain barrier function, and pericyte dysfunction may contribute to the pathophysiology of human neurological diseases including Alzheimers disease, multiple sclerosis, and stroke. Induced pluripotent stem cell (iPSC)-derived pericytes (iPericytes) are a promising tool for vascular research. However, it is unclear how iPericytes functionally compare to primary human brain vascular pericytes (HBVPs). We differentiated iPSCs into iPericytes of either the mesoderm or neural crest lineage using established protocols. We compared iPericyte and HBVP morphologies, quantified gene expression by qPCR and bulk RNA sequencing, and visualised pericyte protein markers by immunocytochemistry. To determine whether the gene expression of neural crest iPericytes, mesoderm iPericytes or HBVPs correlated with their functional characteristicsin vitro, we quantified EdU incorporation following exposure to the key pericyte mitogen, platelet derived growth factor (PDGF)-BB and, contraction and relaxation in response to the vasoconstrictor endothelin-1 or vasodilator adenosine, respectively. iPericytes were morphologically similar to HBVPs and expressed canonical pericyte markers. However, iPericytes had 1864 differentially expressed genes compared to HBVPs, while there were 797 genes differentially expressed between neural crest and mesoderm iPericytes. Consistent with the ability of HBVPs to respond to PDGF-BB signalling, PDGF-BB enhanced and PDGF receptor-beta inhibitors impaired iPericyte proliferation. Administration of endothelin-1 led to iPericyte contraction and adenosine led to iPericyte relaxation, of a magnitude similar to the response evoked in HBVPs. We determined that neural crest iPericytes were less susceptible to PDGFR beta inhibition, but responded most robustly to vasoconstrictive meditators. iPericytes express pericyte-associated genes and proteins and, exhibit an appropriate physiological response upon exposure to a key endogenous mitogen or vasoactive mediators. Therefore, the generation of functional iPericytes would be suitable for use in future investigations exploring pericyte function or dysfunction in neurological diseases.

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“…Remarkably, effective brain repair was observed, characterized by the presence of numerous neurons exhibiting typical neuronal morphology, complete with axons, dendrites, and the ability to generate action potentials [203]. Brain organoids ALS and FTD iPSC Granulin loss of function in human mature brain organoids implicates astrocytes in TDP-43 pathology [229] Motor neurons ALS hiPSC Exploring motor neuron diseases using iPSC platforms [230] Cerebral organoids FTD iPSC ELAVL4, splicing, and glutamatergic dysfunction precede neuron loss in MAPT mutation cerebral organoids [133] Molecular study FTD iPSC Pathological progression induced by the frontotemporal dementia-associated R406W tau mutation in patient-derived iPSCs [231] iPSC-derived astrocytes MS iPSC iPSC-derived reactive astrocytes from patients with multiple sclerosis protect cocultured neurons in inflammatory conditions [232] Model study MS iPSC Selective PDE4 subtype inhibition provides new opportunities to intervene in neuroinflammatory versus myelin-damaging hallmarks of multiple sclerosis [233] RRMS and PPMS iPSC cellular models MS iPSC Generation of RRMS-and PPMS-specific iPSCs as a platform for modeling multiple sclerosis [234] Cerebral organoids MS iPSC Cerebral organoids in primary progressive multiple sclerosis reveal stem cell and oligodendrocyte differentiation defect [165] Model study MS iPSC Generation and characterization of four multiple sclerosis iPSC lines from a single family [235] Cerebral organoids ASD iPSC Single-cell brain organoid screening identifies developmental defects in autism [236] Forebrain organoids/Molecular study…”

Section: Traumatic Brain Injurymentioning

confidence: 99%

Induced Pluripotent Stem Cells and Organoids in Advancing Neuropathology Research and Therapies

Pazzin,

Previato,

Budelon Gonçalves

et al. 2024

Cells

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This review delves into the groundbreaking impact of induced pluripotent stem cells (iPSCs) and three-dimensional organoid models in propelling forward neuropathology research. With a focus on neurodegenerative diseases, neuromotor disorders, and related conditions, iPSCs provide a platform for personalized disease modeling, holding significant potential for regenerative therapy and drug discovery. The adaptability of iPSCs, along with associated methodologies, enables the generation of various types of neural cell differentiations and their integration into three-dimensional organoid models, effectively replicating complex tissue structures in vitro. Key advancements in organoid and iPSC generation protocols, alongside the careful selection of donor cell types, are emphasized as critical steps in harnessing these technologies to mitigate tumorigenic risks and other hurdles. Encouragingly, iPSCs show promising outcomes in regenerative therapies, as evidenced by their successful application in animal models.

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Generation and characterisation of four multiple sclerosis iPSC lines from a single family

Cited by 4 publications

References 3 publications

Induced pluripotent stem cell derived pericytes respond to mediators of proliferation and contractility

Induced pluripotent stem cell derived pericytes respond to mediators of proliferation and contractility

Induced pluripotent stem cell derived pericytes respond to endogenous mediators of proliferation and contractility

Induced Pluripotent Stem Cells and Organoids in Advancing Neuropathology Research and Therapies

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