Secretomes of human pluripotent stem cell-derived smooth muscle cell progenitors upregulate extracellular matrix metabolism in the lower urinary tract and vagina

Abstract

Background Adult mesenchymal stem cells (MSCs) have been studied extensively for regenerative medicine; however, they have limited proliferation in vitro, and the long culture time induces cell senescence. MSCs also contribute to tissue repair through their paracrine function. In this study, we sought to examine the paracrine effects of human smooth muscle cell progenitors (pSMC) on the urethra and adjacent vagina of stress urinary incontinence rodents. We use human pluripotent stem cell (PSC) lines to derive pSMCs to overcome the issue of decreased proliferation in tissue culture and to obtain a homogenous cell population. Method Three human PSC lines were differentiated into pSMCs. The conditioned medium (CM) from pSMC culture, which contain pSMC secretomes, was harvested. To examine the effect of the CM on the extracellular matrix of the lower urinary tract, human bladder smooth muscle cells (bSMCs) and vaginal fibroblasts were treated with pSMC-CM in vitro. Stress urinary incontinence (SUI) was induced in rats by surgical injury of the urethra and adjacent vagina. SUI rats were treated with pSMC-CM and monitored for 5 weeks. Urethral pressure testing was performed prior to euthanasia, and tissues were harvested for PCR, Western blot, and histological staining. Kruskal-Wallis one-way ANOVA test and Student t test were used for statistical comparisons. Results pSMC-CM upregulated MMP-2, TIMP-2, collagen, and elastin gene expression, and MMP-9 activity in the human bladder and vaginal cells consistent with elastin metabolism modulation. pSMC-CM treatment in the SUI rat improved urethral pressure (increase in leak point pressure compared to intact controls, p < 0.05) and increased collagen and elastin expression in the urethra and the adjacent vagina. Conclusion Conditioned media from smooth muscle cell progenitors derived from human pluripotent stem cells improved urethral leak point pressure and collagen and elastin content in the SUI rat. These findings suggest a novel therapeutic potential for PSC-based treatments for SUI and pelvic floor disorders where tissues are affected by collagen, elastin, and smooth muscle loss.

Mesenchymal stem cell-based therapy for female stress urinary incontinence

“…Similarly, Deng et al (2015) demonstrated the ability of MSCs to repair damaged tissue in dual injuries of muscles and nerves via secretion of bioactive factors. Other studies have shown that the feasibility of CCM in SUI treatment is similar to that of MSCs (Jiang et al, 2021;Zhuang et al, 2021).…”

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

“…This group yielded histologic evidence of extracellular remodeling in the native rodent tissue and confirmed restoration of elastic fibers and muscle function after pSMC injections 7 . Moreover, their recent study suggests a potent paracrine effect of media conditioned by human PSC-derived pSMCs on improvement of function and histologic characteristics of the rat urethra and vagina 33,34 . These data suggest a novel therapeutic potential for PSC-based treatments for SUI and pelvic floor disorders where tissues are affected by loss of smooth muscle and ECM components collagen, or elastin.…”

Functional outcome of the anterior vaginal wall in a pelvic surgery injury rat model after treatment with stem cell-derived progenitors of smooth muscle cells

“…Proximal vaginal tissues were embedded in paraffin and proximal urethral tissues were embedded in OCT. Elastin staining on paraffin and OCT were performed as described previously [ 13 , 14 ]. Briefly, elastin fibers were stained in Weigert’s Resorcin-Fuchsin solution (Electron Microscopy Sciences, Hatfield, PA).…”