Mas Aymara, Nair Sangeeta, Laknaur Archana, Simón Carlos, Diamond Michael P, Al-Hendy Ayman
Department of Obstetrics and Gynecology, Georgia Regents University, Augusta, Georgia.
Fundación Instituto Valenciano de Infertilidad, Instituto Universitario IVI-University of Valencia, INCLIVA, Valencia, Spain; Department of Obstetrics and Gynecology, School of Medicine, Stanford University, Stanford, California.
Fertil Steril. 2015 Jul;104(1):225-34.e3. doi: 10.1016/j.fertnstert.2015.04.021. Epub 2015 May 16.
To identify and characterize myometrial/fibroid stem cells by specific stem cell markers in human myometrium, and to better understand the stem cell contribution in the development of uterine fibroids.
Prospective, experimental human and animal study.
University research laboratory.
PATIENT(S)/ANIMAL(S): Women undergoing hysterectomy for treatment of symptomatic uterine fibroids and female NOD/SCID/IL-2Rγ(null) mice.
INTERVENTION(S): Identification and isolation of stem cells from human fibroids and adjacent myometrium tissues using Stro-1/CD44-specific surface markers.
MAIN OUTCOME MEASURE(S): Flow cytometry, semiquantitative polymerase chain reaction, clonogenicity assays, cell culture, molecular analysis, immunocyto-histochemistry, in vitro differentiation, and xenotransplantation assays.
RESULT(S): Using Stro-1/CD44 surface markers, we were able to isolate stem cells from adjacent myometrium and human fibroid tissues. The undifferentiated status of isolated cells was confirmed by the expression of ABCG2 transporter, as well as additional stem cell markers OCT4, NANOG, and GDB3, and the low expression of steroid receptors ERα and PR-A/PR-B. Mesodermal cell origin was established by the presence of typical mesenchymal markers (CD90, CD105, and CD73) and absence of hematopoietic stem cell markers (CD34, CD45), and confirmed by the ability of these cells to differentiate in vitro into adipocytes, osteocytes, and chondrocytes. Finally, their functional capability to form fibroid-like lesions was established in a xenotransplantation mouse model. The injected cells labeled with superparamagnetic iron oxide were tracked by both magnetic resonance imaging and fluorescence imaging, thus demonstrating the regenerative potential of putative fibroid stem cells in vivo.
CONCLUSION(S): We have demonstrated that Stro-1/CD44 can be used as specific surface markers to enrich a subpopulation of myometrial/fibroids cells, exhibiting key features of stem/progenitor cells. These findings offer a useful tool to better understand the initiation of uterine fibroids, and may lead to the establishment of effective therapeutic options.
通过人子宫肌层中特定的干细胞标志物来鉴定和表征子宫肌层/肌瘤干细胞，并更好地理解干细胞在子宫肌瘤发生发展中的作用。
前瞻性实验性人体和动物研究。
大学研究实验室。
患者/动物：因症状性子宫肌瘤接受子宫切除术的女性以及雌性NOD/SCID/IL-2Rγ(null)小鼠。
使用Stro-1/CD44特异性表面标志物从人子宫肌瘤及相邻子宫肌层组织中鉴定和分离干细胞。
流式细胞术、半定量聚合酶链反应、克隆形成试验、细胞培养、分子分析、免疫细胞化学、体外分化和异种移植试验。
利用Stro-1/CD44表面标志物，我们能够从相邻子宫肌层和人子宫肌瘤组织中分离出干细胞。通过ABCG2转运蛋白以及其他干细胞标志物OCT4、NANOG和GDB3的表达，以及类固醇受体ERα和PR-A/PR-B的低表达，证实了分离细胞的未分化状态。通过典型间充质标志物（CD90、CD105和CD73）的存在以及造血干细胞标志物（CD34、CD45）的缺失确定了中胚层细胞起源，并通过这些细胞在体外分化为脂肪细胞、骨细胞和软骨细胞的能力得到证实。最后，在异种移植小鼠模型中确定了它们形成肌瘤样病变的功能能力。通过磁共振成像和荧光成像追踪注射了超顺磁性氧化铁标记的细胞，从而证明了假定的肌瘤干细胞在体内的再生潜力。
我们已经证明Stro-1/CD44可作为特异性表面标志物来富集子宫肌层/肌瘤细胞亚群，这些细胞表现出干/祖细胞的关键特征。这些发现为更好地理解子宫肌瘤的发病机制提供了有用的工具，并可能有助于建立有效的治疗方案。
