Therapeutic Efficacy of Human Embryonic Stem Cell-Derived Multipotent Stem/Stromal Cells in Diabetic Detrusor Underactivity: A Preclinical Study

Abstract

Mesenchymal stem/stromal cell (MSC) therapy is a promising approach for treatment of as yet incurable detrusor underactivity (DUA), which is characterized by decreased detrusor contraction strength and/or duration, leading to prolonged bladder emptying. In the present study, we demonstrated the therapeutic potential of human embryonic stem cell (ESC)-derived multipotent MSCs (M-MSCs) in a diabetic rat model of DUA. Diabetes mellitus (DM) was induced by intraperitoneal injection of streptozotocin (STZ) (50 mg/kg) into 8-week-old female Sprague-Dawley rats. Three weeks later, various doses of M-MSCs (0.25, 0.5, and 1 × 106 cells) or an equivalent volume of PBS were injected into the outer layer of the bladder. Awake cystometry, organ bath, histological, and gene expression analyses were evaluated 1 week (short-term) or 2 and 4 weeks (long-term) after M-MSC transplantation. STZ-induced diabetic rats developed DUA, including phenotypes with significantly longer micturition intervals, increased residual urine amounts and bladder capacity, decreased micturition pressure on awake cystometry, and contractile responses to various stimuli in organ bath studies. Muscle degeneration, mast cell infiltration, fibrosis, and apoptosis were present in the bladders of DM animals. A single local transplantation of M-MSCs ameliorated DUA bladder pathology, including functional changes and histological evaluation, and caused few adverse outcomes. Immunostaining and gene expression analysis revealed that the transplanted M-MSCs supported myogenic restoration primarily by engrafting into bladder tissue via pericytes, and subsequently exerting paracrine effects to prevent apoptotic cell death in bladder tissue. The therapeutic efficacy of M-MSCs was superior to that of human umbilical cord-derived MSCs at the early time point (1 week). However, the difference in efficacy between M-MSCs and human umbilical cord-derived MSCs was statistically insignificant at the later time points (2 and 4 weeks). Collectively, the present study provides the first evidence for improved therapeutic efficacy of a human ESC derivative in a preclinical model of DM-associated DUA.

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

“…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)].…”

“…Detrusor pressure was defined as [intravesical pressure (IVP)intraabdominal pressure (IAP)]. The contractility of the bladder tissues was measured by the organ bath study, as previously described [12,19]. In brief, longitudinal strips of posterior bladder wall were normalized to weight per 1 g and then mounted in 5 mL organ baths containing Krebs solution (118 mM NaCl, 5.0 mM KCl, 2.5 mM CaCl 2 , 1.0 mM MgSO 4 , 30 mM NaHCO 3 , 1.0 mM KH2PO 4 , and 11.4 mM glucose, pH 7.4), and maintained at 37°C with 5% CO 2 and 95% O 2 continuously supplied.…”

“…In brief, longitudinal strips of posterior bladder wall were normalized to weight per 1 g and then mounted in 5 mL organ baths containing Krebs solution (118 mM NaCl, 5.0 mM KCl, 2.5 mM CaCl 2 , 1.0 mM MgSO 4 , 30 mM NaHCO 3 , 1.0 mM KH2PO 4 , and 11.4 mM glucose, pH 7.4), and maintained at 37°C with 5% CO 2 and 95% O 2 continuously supplied. The contractile response to 80 mM KCl (P9333; Sigma-Aldrich, St. Louis, MO, USA), EFS (electrical field stimulation, 1, 2, 4, 8, 16, and 32 Hz), 1 mM ATP (A2383; Sigma-Aldrich), or carbachol concentration (PHR1511; Sigma-Aldrich; 1 nM to 1 mM) were recorded as previously described [12,19]. An electrical pulse (1 millisecond pulse width and 80 V in bath) was delivered using a stimulator (D-7806; Hugo Sachs Elektronik, Germany) for 5 s at increasing frequencies (1,2,4,8,16, and 32 Hz), with 5 min intervals between electrical field stimulations.…”

“…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].…”

“…Previously, we investigated the therapeutic effects of multipotent MSCs (M-MSCs) derived from human ESCs in a streptozotocin-induced diabetic rat model of DUA. The transplanted M-MSCs integrated into pericytes, which provided favorable micro-environments for detrusor muscle regeneration [12]. In the present study, we aimed to investigate the therapeutic effects of M-MSC treatment in a DUA rat model with extensive vascular endothelial damage of the iliac arteries.…”