The use of placental mesenchymal stem cells to repair the damaged endometrium

doi:10.3969/j.issn.2095-4344.2017.13.018

Abstract

Abstract:

BACKGROUND: Placental mesenchymal stem cells are rich in source and easily obtained, which can differentiate into osteoblasts, nerve cells and liver cells. Additionally, there is no immune rejection and ethical issues in the clinical application. Therefore, placental mesenchymal stem cells are considered to be a good source of adult stem cells.
OBJECTIVE: To observe the effect of placental mesenchymal stem cell transplantation in the repair of endometrial lesions in rats.
METHODS: Endometrial damage models were established in rats by means of thermal damage, and on the 15th day after modeling, these rat models were randomly divided into four groups (n=10 per group): intrauterine injection of 1 mL of allogeneic placenta mesenchymal stem cell suspension (intrauterine transplantation group), intrauterine injection of the same amount of PBS (intrauterine control group), tail vein injection of 1 mL of allogeneic placenta mesenchymal stem cell suspension (intravenous transplantation group), and tail vein injection of the same amount of PBS (intravenous control group). The female rats experiencing the third estrus after modeling were caged with male rats to observe whether the vaginal plug appeared. The female rats were killed the same day when the vaginal plug was observed, and uterus tissues were taken to detect the number of endometrial glands as well as perform immunohistochemistry and western blot detection.
RESULTS AND CONCLUSION: The number of endometrial glands was highest in the intrauterine transplantation group followed by the intravenous transplantation group, and lowest in the two control groups (P < 0.05). The expression of integrin ανβ3 shown by immunohistochemistry and western blot was highest in the intrauterine transplantation group followed by the intravenous transplantation, and lowest in the two control groups (P < 0.05). These findings indicate that the placental mesenchymal stem cell transplantation can repair damaged endometrial tissues in rats to different degrees, by increasing endometrial glands count and improving the endometrial receptivity.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cell Transplantation, Endometrium, Placenta, Integrin alphaVbeta3, Tissue Engineering

CLC Number:

R394.2

Niu Ting, Li Ai-bin, Chen Li. The use of placental mesenchymal stem cells to repair the damaged endometrium[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(13): 2074-2080.

Figures/Tables 3

2.1 大鼠子宫内膜损伤造模情况 2.1.1 大鼠正常子宫大体解剖及组织病理切片苏木精-伊红染色显微镜下观察大鼠正常子宫内膜组织，可见宫腔形态不规则，分为内膜层、肌层和外膜层，子宫内膜呈单层柱状上皮，覆盖子宫腔表面和腺腔，子宫内膜腺体主要位于黏膜下层和基底层，呈椭圆形或圆形(图1A)。 2.1.2 热损伤处理后大鼠子宫内膜组织建模后1 d，大体可见子宫充血、肿大，苏木精-伊红染色显微镜下观察可见内膜间质充血、水肿，毛细血管扩张，其内可见大量红细胞；处理后2 d，大体见子宫水肿减轻，苏木精-伊红染色镜下可见内膜间质仍有血管充血，宫腔内可见脱落的子宫内膜组织碎片，子宫内膜组织结构模糊、紊乱；处理后7 d，大体可见受损子宫与周围组织粘连，苏木精-伊红染色镜下可见子宫内膜层变薄，腺体萎缩、变小，部分宫腔腔上皮内膜处中断；处理后14 d，大体见子宫形态缩小、苍白，苏木精-伊红染色见子宫腔变小，正常的内膜组织较少, 部分组织开始纤维化，苏木精-伊红染色显示为较多红色组织(图1B-E)。 2.1.3 生育能力检测造模后第15天，将造模大鼠与雄性鼠合笼，观察到大鼠阴栓，记录为孕0.5 d，雌鼠受孕12 d 后处死，可见处理侧子宫萎缩变小，外观显示为白色条索状，子宫胚鼠数目为一两个，而对侧正常子宫侧可见胚鼠6-8只(图2)；统计学分析结果显示：内膜损伤侧子宫与正常子宫侧胚泡数目比较差异有显著性意义(1.00±0.82，6.00±0.97，P < 0.05)。 2.2 细胞移植后修复效果检测 2.2.1 细胞移植后大鼠子宫内膜腺体计数细胞移植后，通过阴道脱落细胞涂片法确定大鼠动情周期，在第3个动情周期的动情期断颈处死大鼠，对各组横切面子宫内膜腺体计数，结果见表1，其中宫腔移植组子宫内膜腺体计数最多，尾静脉移植组腺体个数稍少，单因素方差分析检验各组子宫内膜腺体计数，各组间差异有显著性意义(P < 0.05)；SNK检验显示，宫腔移植组腺体个数多于宫腔对照组、尾静脉移植组、尾静脉对照组(P < 0.05)，尾静脉移植组多于宫腔对照组、尾静脉对照组(P < 0.05)，宫腔对照组与尾静脉对照组间无差异。 2.2.2 免疫组织化学检测子宫内膜整合素ανβ3表达见图3。 ανβ3表达于子宫内膜腺上皮和腔上皮，是子宫内膜容受性的标记之一，表达于大鼠围着床期，可用于监测子宫内膜损伤修复的指标之一，用IPP软件来计算各组间ανβ3的吸光度值，结果见表1，宫腔移植组ανβ3的表达最强，尾静脉移植组稍弱，单因素方差分析检验各组间差异有显著性意义(P < 0.05)；SNK检验显示，宫腔移植组整合素ανβ3表达强于宫腔对照组、尾静脉移植组、尾静脉对照组(P < 0.05)，尾静脉移植组强于宫腔对照组、尾静脉对照组(P < 0.05)，宫腔对照组与尾静脉对照组间无差异。 2.2.3 Western blot检测子宫内膜整合素ανβ3的表达见图4。宫腔移植组ανβ3表达最强，尾静脉移植组稍弱，见表1，单因素方差分析检验各组间差异有显著性意义(P < 0.05)；SNK检验显示，宫腔移植组整合素ανβ3表达强于宫腔对照组、尾静脉移植组、尾静脉对照组(P < 0.05)，尾静脉移植组强于宫腔对照组、尾静脉对照组(P < 0.05)，宫腔对照组与尾静脉对照组间无差异。"

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