Stimulating vaginal repair in rats through skeletal muscle-derived stem cells seeded on small intestinal submucosal scaffolds.
Stimulating vaginal repair in rats through skeletal muscle-derived stem cells seeded on small intestinal submucosal scaffolds.
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DOI：
10.1097/aog.0b013e3181af6abd
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发表时间：
2009-08
影响因子：
7.2
通讯作者：
Gonzalez-Cadavid NF
中科院分区：
文献类型：
--
作者：
Ho MH;Heydarkhan S;Vernet D;Kovanecz I;Ferrini MG;Bhatia NN;Gonzalez-Cadavid NF
移植物用于脱垂后的阴道修复，但它们用于携带干细胞再生阴道组织的报道还没有。在这项研究中，我们研究了1）在小肠粘膜下层（SIS）上生长的肌源性干细胞（MDSC）是否在体外和植入阴道缺陷大鼠模型后产生平滑肌细胞（SMC）; 2）表达适用于阴道内源性干细胞体内检测的标志物; 3）刺激阴道修复。通过免疫细胞化学、蛋白质印迹和实时聚合酶链反应（PCR）检测在单层、SIS或聚合物网状物上生长的小鼠MDSC的细胞分化。干细胞标记物通过DNA微阵列进行筛选，然后进行实时PCR、免疫细胞化学和Western印迹。将经历子宫切除术和部分阴道切除术的大鼠原样保留或在阴道中植入SIS上的4 '，6-二脒基-2-苯基吲哚（DAPI）标记的MDSC或无MDSC的SIS，免疫抑制，并在2-8周时处死。将免疫荧光、苏木精-伊红和Masson三色应用于组织切片。单层和支架上的肌源性干细胞培养物分化为SMC，如α-平滑肌肌动蛋白（ASMA）、钙调蛋白和平滑肌蛋白标记物所示。肌源性干细胞表达胚胎干细胞标志物Oct-4和nanog。双重DAPI/ASMA荧光指示MDSC转化为SMC。肌肉来源的干细胞/SIS刺激阴道组织修复，包括角蛋白-5阳性上皮形成，并在4周和8周时防止纤维化。鉴定了Oct-4+推定的内源性干细胞。肌源性干细胞/SIS植入物刺激大鼠阴道组织修复，因此支架上的自体MDSC可能是治疗阴道修复的有前途的方法。
Grafts are used for vaginal repair after prolapse, but their use to carry stem cells to regenerate vaginal tissue has not been reported. In this study, we investigated whether 1) muscle-derived stem cells (MDSC) grown on small intestinal submucosa (SIS) generate smooth-muscle cells (SMC) in vitro and upon implantation in a rat model of vaginal defects; 2) express markers applicable to the in-vivo detection of vaginal endogenous stem cells; and 3) stimulate the repair of the vagina. Mouse MDSC grown on monolayer, SIS, or polymeric mesh, were tested for cell differentiation by immunocytochemistry, Western blot and real-time polymerase chain reaction (PCR). Stem cell markers were screened by DNA microarrays followed by real-time PCR, immunocytochemistry, and Western blot. Rats that underwent hysterectomy and partial vaginectomy were left as such or implanted in the vagina with 4’,6-Diamidino-2-Phenylindole (DAPI)–labeled MDSC on SIS, or SIS without MDSC, immunosuppressed, and killed at 2–8 weeks. Immunofluorescence, hematoxylin-eosin, and Masson trichrome were applied to tissue sections. Muscle-derived stem cell cultures on monolayer and on scaffolds differentiate into SMC, as shown by α-smooth muscle actin (ASMA), calponin, and smoothelin markers. Muscle-derived stem cells express embryonic stem cell markers Oct-4 and nanog. Dual DAPI/ASMA fluorescence indicated MDSC conversion to SMC. Muscle-derived stem cells/SIS stimulated vaginal tissue repair, including keratin-5 positive epithelium formation and prevented fibrosis at 4 and 8 weeks. Oct-4+ putative endogenous stem cells were identified. Muscle-derived stem cells/SIS implants stimulate vaginal tissue repair in the rat, thus autologous MDSC on scaffolds may be a promising approach for the treatment of vaginal repair.