标记滞留细胞技术结合干细胞标记物检测小鼠子宫内膜干细胞

目的通过标记滞留细胞技术检测昆明小鼠子宫内膜干细胞的存在及其分布情况;观察P63和Musashi-1在不同周龄小鼠子宫内膜的表达以及两者与标记滞留细胞的关系,探讨P63和Musashi-1作为子宫内膜干细胞特异性标记物的可能性。方法出生3天雌性昆明小鼠皮下注射BrdU,分别在1w、2w、3w、4w、6w、8w和10w处死小鼠取其子宫。采用免疫组化法分别检测BrdU、P63和Musashi-1在各周龄小鼠子宫内膜的表达情况。结果标记后1w的小鼠,子宫内膜绝大部分的上皮和基质细胞都被BrdU标记。随着小鼠周龄的增加,子宫内膜BrdU阳性细胞百分率逐渐降低。至第8w时,仅在基质中有极少量的BrdU阳性细胞,主要位于基质与肌层交界处。早期小鼠子宫内膜组织中P63和Musashi-1阳性细胞数量较多,随着子宫内膜的逐渐发育成熟,P63和Musashi-1的表达逐渐减少,其表达规律及分布与标记滞留细胞基本一致。结论 (1)小鼠子宫内膜标记滞留细胞主要位于基质内膜与肌层交界处,这些细胞的分布与推测的子宫内膜干细胞的分布部位相符。(2)P63和Musashi-1是较特异的干细胞标记物。

Using the label-retaining cell technique and stem cell markers to detect endometrial stem cells

Objective To observe the endometrial stem cells in mice by using label-retaining cell （LRC） technique, and to ascertain whether P63 and Musashi-1 could be used as specific stem cell markers. Methods 3-day-old white Kunming mice were subcutaneously injected with BrdU twice daily for 3 consecutive days. They were sacrificed to harvest their uteri at weeks 1, 2, 3, 4, 6, 8 and 10. Immunohistochemistry was used to analyze BrdU, P63 and Musashi-l＂expressions in the endometrium. Results At I week af- ter BrdU injection, the majority of epithelial and stromal cells were stained for BrdU. The percentage of BrdU-positive cells began to decrease gradually thereafter. After the 8th week, almost no BrdU-positive cells were found in the endometrial epithelium, but a few stromal cells were BrdU-positive, and most of them were located at the interface of stromal and myornetrial junction. The P63 and Musashi-1 positive cells were more numerous in the early development, but with the development and maturation, the positive cells began to decrease gradually. The expression of P63 and Musashi-1 accorded with that of BrdU. Conclusion （1） LRCs are located mainly at the stroma in the interface of endometrial -myometrial junction. （2） P63 and Musashi-1 may be used as assistant endometrial stem cell markers.