Differentiation of patient-specific void urine-derived human induced pluripotent stem cells to fibroblasts and skeletal muscle myocytes

Cell-based therapy is a major focus for treatment of stress urinary incontinence (SUI). However, derivation of primary cells requires tissue biopsies, which often have adverse effects on patients. A recent study used human induced pluripotent stem cells (iPSC)-derived smooth muscle myocytes for uret...

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Bibliographic Details
Published in:Scientific reports 2023-03, Vol.13 (1), p.4746-4746, Article 4746
Main Authors: Kibschull, M., Nguyen, T. T. N., Chow, T., Alarab, M., Lye, S. J., Rogers, I., Shynlova, O.
Format: Article
Language:English
Subjects:
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631/80
692/4025
Animal models
Animals
Biopsy
Blue stain
Cell culture
Cell Differentiation
Cells, Cultured
Connectin
Desmin
Female
Fibroblasts
Fibroblasts - metabolism
Fibronectin
Flow cytometry
Humanities and Social Sciences
Humans
Induced Pluripotent Stem Cells
Mitochondria
multidisciplinary
Muscle Fibers, Skeletal
Muscle, Skeletal
Myocytes
MyoD protein
Nanoparticles
Paxillin
Pluripotency
Rats
Science
Science (multidisciplinary)
Skeletal muscle
Smooth muscle
Sphincter
Stem cells
Urinary incontinence
Urinary Incontinence, Stress - therapy
Vimentin
Vinculin
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Summary:Cell-based therapy is a major focus for treatment of stress urinary incontinence (SUI). However, derivation of primary cells requires tissue biopsies, which often have adverse effects on patients. A recent study used human induced pluripotent stem cells (iPSC)-derived smooth muscle myocytes for urethral sphincter regeneration in rats. Here, we establish a workflow using iPSC-derived fibroblasts and skeletal myocytes for urethral tissue regeneration: (1) Cells from voided urine of women were reprogrammed into iPSC. (2) The iPSC line U1 and hESC line H9 (control) were differentiated into fibroblasts expressing FSP1, TE7, vinculin, vimentin, αSMA, fibronectin and paxillin. (3) Myogenic differentiation of U1 and H9 was induced by small molecule CHIR99021 and confirmed by protein expression of myogenic factors PAX7, MYOD, MYOG, and MF20. Striated muscle cells enriched by FACS expressed NCAM1, TITIN, DESMIN, TNNT3. (4) Human iPSC-derived fibroblasts and myocytes were engrafted into the periurethral region of RNU rats. Injected cells were labelled with ferric nanoparticles and traced by Prussian Blue stain, human-specific nuclear protein KU80, and human anti-mitochondria antibody. This workflow allows the scalable derivation, culture, and in vivo tracing of patient-specific fibroblasts and myocytes, which can be assessed in rat SUI models to regenerate urethral damages and restore continence.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-023-31780-9