Longitudinal intravital imaging of transplanted mesenchymal stem cells elucidates their functional integration and therapeutic potency in an animal model of interstitial cystitis/bladder pain syndrome

Abstract

Rationale: Mesenchymal stem cell (MSC) therapy may be a novel approach to improve interstitial cystitis/bladder pain syndrome (IC/BPS), an intractable disease characterized by severe pelvic pain and urinary frequency. Unfortunately, the properties of transplanted stem cells have not been directly analyzed in vivo, which hampers elucidation of the therapeutic mechanisms of these cells and optimization of transplantation protocols. Here, we monitored the behaviors of multipotent stem cells (M-MSCs) derived from human embryonic stem cells (hESCs) in real time using a novel combination of in vivo confocal endoscopic and microscopic imaging and demonstrated their improved therapeutic potency in a chronic IC/BPS animal model. Methods: Ten-week-old female Sprague-Dawley rats were instilled with 10 mg of protamine sulfate followed by 750 µg of lipopolysaccharide weekly for 5 weeks. The sham group was instilled with phosphate-buffered saline (PBS). Thereafter, the indicated dose (0.1, 0.25, 0.5, and 1×10 6 cells) of M-MSCs or PBS was injected once into the outer layer of the bladder. The distribution, perivascular integration, and therapeutic effects of M-MSCs were monitored by in vivo endoscopic and confocal microscopic imaging, awake cystometry, and histological and gene expression analyses. Results: A novel combination of longitudinal intravital confocal fluorescence imaging and microcystoscopy in living animals, together with immunofluorescence analysis of bladder tissues, demonstrated that transplanted M-MSCs engrafted following differentiation into multiple cell types and gradually integrated into a perivascular-like structure until 30 days after transplantation. The beneficial effects of transplanted M-MSCs on bladder voiding function and the pathological characteristics of the bladder were efficient and long-lasting due to the stable engraftment of these cells.

A Preclinical Study of Human Embryonic Stem Cell-Derived Mesenchymal Stem Cells for Treating Detrusor Underactivity by Chronic Bladder Ischemia

“…1). In addition, M-MSCs up-regulated their expression of WNT family member genes (e.g., WNT2, WNT4, and WNT5B), which play an important role in repair of the bladder injury [9,10,14,24]. Taken together, these results support the finding that M-MSCs, mainly engrafted as pericytes in blood vessels near bladder muscle fibers, stimulate angiogenesis in the CBI injured bladder, which might be crucial for their therapeutic potency.…”

“…Additional rats (n = 6) were used for the organ bath study. The M-MSCs used in the stem cell treatments were maintained up to seven passages only to ensure their functionality as previously described [9,10,18].…”

“…One week after M-MSC injection, the experimental groups were evaluated by awake cystometry. Bladder voiding function was evaluated in unrestrained, awake-state rats in metabolic cages as previously described [9,10,12]. Detrusor pressure was defined as [intravesical pressure (IVP)intraabdominal pressure (IAP)].…”

“…MSCs can been isolated from adult or fetal tissues [5], or they can be also derived from pluripotent stem cells (PSCs) such as embryonic stem cells (ESCs) and induced PSCs (iPSCs) [6][7][8]. The beneficial effects of MSC therapy include the generation of various preclinical bladder dysfunction models and reports of positive patient outcomes from clinical trials [9][10][11][12][13][14][15][16][17].…”

Manipulated Mesenchymal Stem Cells Applications in Neurodegenerative Diseases

“…The PSCs derived-MSCs replace damaged cells in injured tissues, exert immunomodulatory effects, release matrix proteins and growth factors that eventually adjusts the surroundings of damaged tissues. These PSCs-MSCs are evidently useful in regenerative medicine to treat a number of unmanageable immunological and neurological disorders (5,6).…”