Generation of functional endothelial cells with progenitor-like features from murine induced pluripotent stem cells

Kachamakova‐Trojanowska, Neli; Nowak, Witold; Szade, Krzysztof; Stępniewski, Jacek; Bukowska-Straková, Karolina; Żukowska, Monika; Taha, Hevidar; Chmura-Skirlińska, Antonina; Beilharz, Michael; Dulak, Józef; Józkowicz, Alicja

doi:10.1016/j.vph.2016.07.008

Cited by 5 publications

(3 citation statements)

References 79 publications

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“…For detection of intracellular as well as extracellular proteins, hiPSCs and iPSC-ECs were first seeded on culture slides (BD Biosciences, Franklin Lakes, NJ, USA), then cultured for 24–72 h in appropriate culture medium. The staining procedure was performed as described previously [18] with the only change of TBS to PBS. All primary and secondary antibodies used in the current study are listed in Table 1.…”

Section: Methodsmentioning

confidence: 99%

Human iPSCs-Derived Endothelial Cells with Mutation in HNF1A as a Model of Maturity-Onset Diabetes of the Young

Kachamakova‐Trojanowska

Stępniewski

Dulak

2019

Cells

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Patients with HNF1A-maturity-onset diabetes of the young (MODY) often develop endothelial dysfunction and related microvascular complications, like retinopathy. As the clinical phenotype of HNF1A-MODY diabetes varies considerably, we used human induced pluripotent stem cells (hiPSCs) from two healthy individuals (control) to generate isogenic lines with mutation in HNF1A gene. Subsequently, control hiPSCs and their respective HNF1A clones were differentiated toward endothelial cells (hiPSC-ECs) and different markers/functions were compared. Human iPSC-ECs from all cell lines showed similar expression of CD31 and Tie-2. VE-cadherin expression was lower in HNF1A-mutated isogenic lines, but only in clones derived from one control hiPSCs. In the other isogenic set and cells derived from HNF1A-MODY patients, no difference in VE-cadherin expression was observed, suggesting the impact of the genetic background on this endothelial marker. All tested hiPSC-ECs showed an expected angiogenic response regardless of the mutation introduced. Isogenic hiPSC-ECs responded similarly to stimulation with pro-inflammatory cytokine TNF-α with the increase in ICAM-1 and permeability, however, HNF1A mutated hiPSC-ECs showed higher permeability in comparison to the control cells. Summarizing, both mono- and biallelic mutations of HNF1A in hiPSC-ECs lead to increased permeability in response to TNF-α in normal glycemic conditions, which may have relevance to HNF1A-MODY microvascular complications.

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Section: Methodsmentioning

confidence: 99%

Human iPSCs-Derived Endothelial Cells with Mutation in HNF1A as a Model of Maturity-Onset Diabetes of the Young

Kachamakova‐Trojanowska

Stępniewski

Dulak

2019

Cells

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“…The final paper in the special issue is an original study by Kachamakova-Trojanowska et al [13] in which the angiogenic capacity of functional murine iPSC-derived ECs are directly compared to bone marrow derived mesenchymal stromal cells (MSCs), which are claimed to possess similar features. The study demonstrates that, in contrast to iPSC-derived ECs, murine MSCs do not reveal efficient blood vesselforming capacities.…”

Section: Editorialmentioning

confidence: 99%

Vascular biology: New mechanisms and pathways

Dulak

Alexander

Randi

2016

Vascular Pharmacology

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“…It has been demonstrated that iPSCs are capable of differentiation into vascular cell phenotypes in response to de ned media and culture conditions providing an autologous and abundant source for application in these elds (26). A range of protocols for differentiating murine and human iPSCs into functional ECs (27,28) and vascular VSMCs (29,30) have been published, with ongoing efforts to further delineate more e cient methods to obtain an increasing yield of proliferative mature vascular cells.…”

Section: Introductionmentioning

confidence: 99%

Vascular cells differentiated from peripheral blood mononuclear cell- versus urine cell-derived induced pluripotent stem cells: A comparative analysis

Deinsberger,

Holzner,

Bromberger

et al. 2023

Preprint

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Background: The use of peripheral blood mononuclear cells (PBMCs) and urine-derived epithelial cells for reprogramming towards induced pluripotent stem cells (iPSCs) has shown to be highly effective. Due to their easy accessibility, these cell sources hold promising potential for non-invasive and repetitive isolation from patients. This study aims to conduct a comparative analysis of the phenotype, differentiation efficacy, and functional properties of iPSCs derived from PBMCs and urine towards endothelial cells (EC) and vascular smooth muscle cells (VSMC). Methods: PBMC-derived iPSCs and urine-derived iPSCs were differentiated to ECs via embryoid body formation, followed by an in-vitro monolayer culture. SMCs were generated through a defined monolayer culture. The expression profiles of iPSCs, iPSC-derived ECs, and iPSC-derived VSMCs were assessed through various techniques such as immunofluorescence, RT-qPCR, western blot, and flow cytometry analysis. Functionality of ECs was evaluated with a tube formation assay, while the functional properties of VSMCs were assessed by measuring the contractile response to carbachol. Results: Both PBMC-derived and urine-derived iPSCs were successfully and efficiently differentiated into functional ECs and VSMCs. The efficacy of EC differentiation did not differ significantly between the two cell types, with both yielding approximately 45% mature ECs. The derived ECs displayed morphological and functional characteristics consistent with native ECs, including marker expression and tube formation. However, pluripotency marker SOX2 continued to be upregulated, while OCT4, KLF4, c-Myc, and SSEA-4 were downregulated. Functional assessment via tube formation assays showed no significant difference in the amount of newly formed tubes and branches between the two cell types. VSMC differentiation resulted in 96% and 94% α-SMA positive cells for PBMC-derived and urine-derived iPSCs, respectively. VSMCs of both origins exhibited a spindle-shaped, contractile morphology and expressed α-SMA, calponin, and transgelin consistent with native VSMCs. The generated VSMC lines from both cell sources demonstrated adequate contractility in response to carbachol. Conclusions: This study demonstrates a comparative analysis of functional ECs and VSMCs generated from PBMC-derived and urine-derived iPSCs. Comparison of morphology, expression profile, and functionality of vascular cells generated from both cell sources did not reveal significant differences.

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Generation of functional endothelial cells with progenitor-like features from murine induced pluripotent stem cells

Cited by 5 publications

References 79 publications

Human iPSCs-Derived Endothelial Cells with Mutation in HNF1A as a Model of Maturity-Onset Diabetes of the Young

Human iPSCs-Derived Endothelial Cells with Mutation in HNF1A as a Model of Maturity-Onset Diabetes of the Young

Vascular biology: New mechanisms and pathways

Vascular cells differentiated from peripheral blood mononuclear cell- versus urine cell-derived induced pluripotent stem cells: A comparative analysis

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