Department of Medical and Surgical Sciences, University of Bologna, Bologna 40126, Italy.
Department of Veterinary Medical Sciences, University of Bologna, Bologna 40064, Italy.
World J Gastroenterol. 2019 Sep 7;25(33):4892-4903. doi: 10.3748/wjg.v25.i33.4892.
Mesenchymal stromal cell (MSC)-based therapy is currently under study to treat inflammatory bowel diseases. MSC bioactive products could represent a valid alternative to overcome issues associated with systemic whole-cell therapies. However, MSC anti-inflammatory mechanisms differ between rodents and humans, impairing the reliability of preclinical models.
To evaluate the effect of conditioned medium (CM) derived from porcine vascular wall MSCs (pVW-MSCs) on survival and differentiation of porcine and guinea pig enteric ganglia exposed to lipopolysaccharide (LPS).
Primary cultures of enteric ganglia were obtained by mechanic and enzymatic digestion of ileum resections from guinea pigs () (GPEG) and pigs () (PEG). pVW-MSCs were derived by enzymatic digestion from vascular wall resections of porcine aorta and tested by immunoflowcytometry for MSC immune profile. Enteric ganglia were treated with increasing concentrations of LPS, CM derived by pVW-MSCs or a combination of CM and LPS 1 µg/mL. Cell count and morphometric analysis of HuD positive neurons and glial fibrillary acidic protein positive glial cells were performed by immunofluorecent staining of cultured ganglia.
PEG showed a higher number of neurons compared to GPEG. Overall, CM exerted a protective role on LPS-treated enteric ganglia. CM in combination with LPS increased the number of glial cells per ganglion in both cultures evoking glial cells differentiation in porcine cultures.
These findings suggest an immunomodulating activity of pVW-MSCs mediators on the enteric nervous system in inflammatory conditions.
间充质基质细胞(MSC)为基础的治疗方法目前正在研究用于治疗炎症性肠病。MSC 生物活性产物可能代表一种有效的替代方法,以克服与全身全细胞治疗相关的问题。然而,鼠和人之间 MSC 的抗炎机制不同,这影响了临床前模型的可靠性。
评估猪血管壁间充质基质细胞(pVW-MSCs)来源的条件培养基(CM)对脂多糖(LPS)暴露的猪和豚鼠肠神经节存活和分化的影响。
通过机械和酶消化猪回肠切除术()(PEG)和豚鼠()(GPEG)获得肠神经节的原代培养物。通过酶消化从猪主动脉血管壁获得 pVW-MSCs,并通过免疫流式细胞术测试 MSC 免疫特征。用 LPS、pVW-MSCs 衍生的 CM 或 CM 和 LPS(1 µg/mL)的混合物处理肠神经节。通过培养神经节的免疫荧光染色,对 HuD 阳性神经元和神经胶质纤维酸性蛋白阳性神经胶质细胞的细胞计数和形态计量分析。
PEG 中神经元的数量比 GPEG 多。总的来说,CM 对 LPS 处理的肠神经节发挥了保护作用。CM 与 LPS 联合使用增加了两种培养物中每个神经节的神经胶质细胞数量,在猪培养物中诱导神经胶质细胞分化。
这些发现表明 pVW-MSCs 介质在炎症条件下对肠神经系统具有免疫调节活性。