Department of Obstetrics/Gynecology, Stanford University School of Medicine, 300 Pasteur Drive HH-333, Stanford, CA, 94305, USA.
Department of Obstetrics/Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China.
Stem Cell Res Ther. 2021 Apr 6;12(1):228. doi: 10.1186/s13287-021-02292-y.
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.
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.
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.
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.
成人间充质干细胞（MSCs）已被广泛研究用于再生医学；然而，它们在体外的增殖能力有限，长时间的培养会导致细胞衰老。MSCs 还通过旁分泌功能促进组织修复。在这项研究中，我们试图研究人平滑肌细胞祖细胞（pSMC）对压力性尿失禁啮齿动物尿道和邻近阴道的旁分泌作用。我们使用人多能干细胞（PSC）系来衍生 pSMC，以克服组织培养中增殖能力下降的问题，并获得同质的细胞群体。
将 3 个人 PSC 系分化为 pSMC。收集 pSMC 培养的条件培养基（CM），其中包含 pSMC 分泌组。为了研究 CM 对下尿路细胞外基质的影响，将人膀胱平滑肌细胞（bSMC）和阴道成纤维细胞用 pSMC-CM 进行体外处理。通过尿道和邻近阴道的手术损伤诱导压力性尿失禁（SUI）大鼠。用 pSMC-CM 治疗 SUI 大鼠，并监测 5 周。在安乐死前进行尿道压力测试，并采集组织进行 PCR、Western blot 和组织学染色。使用 Kruskal-Wallis 单向方差分析检验和学生 t 检验进行统计学比较。
pSMC-CM 上调了 MMP-2、TIMP-2、胶原和弹性蛋白基因表达，以及 MMP-9 活性，在人膀胱和阴道细胞中一致，表明弹性蛋白代谢调节。在 SUI 大鼠中，pSMC-CM 治疗可改善尿道压力（与完整对照组相比，漏点压力增加，p<0.05），并增加尿道和邻近阴道的胶原和弹性蛋白表达。
源自人多能干细胞的平滑肌细胞祖细胞的条件培养基可改善 SUI 大鼠的尿道漏点压力和胶原及弹性蛋白含量。这些发现表明，基于 PSC 的治疗方法具有治疗 SUI 和盆底功能障碍的新的治疗潜力，这些疾病的组织受到胶原、弹性蛋白和平滑肌丧失的影响。
