Michalak-Micka Katarzyna, Huerlimann Nadine, Ochsenbein-Kölble Nicole, Ehrbar Martin, Klar Agnes Silvia, Biedermann Thomas, Moehrlen Ueli
Tissue Biology Research Unit, Department of Surgery, University Children's Hospital Zurich, Steinwiesstrasse 75, Zurich, 8032, Switzerland.
Children's Research Center (CRC), University Children's Hospital Zurich, Zurich, Switzerland.
Sci Rep. 2025 Jun 6;15(1):20008. doi: 10.1038/s41598-025-03518-2.
Myelomeningocele (MMC) and myeloschisis (MS) are considered as the most severe forms of spina bifida aperta characterized by incomplete closure of the neural tube during the first trimester of pregnancy. Open maternal-fetal surgery has been identified as a promising alternative for the repair of MMC/MS. The main goal of this study was the isolation and characterization of stem cells derived from MMC/MS amniotic fluid samples. Human amniotic fluid samples were obtained from pregnant women who underwent surgery for fetal spina bifida repair. We applied fluorescence activated-cell sorting (FACS) to immunoselect stem cells from heterogeneous populations of amniocytes based on cKIT (CD117) expression. The cKIT amniocytes were then contrasted with cKIT and unselected amniocytes in order to characterize the phenotype of these cells. cKIT-expressing amniocytes exhibited spindle-shape morphology, while amniocytes negative for cKIT were round-shaped. qRT-PCR analysis revealed significant upregulation of mean mRNA levels of KIT and CD90 genes and downregulation of CK8 gene in cKIT-expressing amniocytes, compared to the negative and unsorted cells. The expression of pluripotency antigens was comparable in sorted cKIT and cKIT amniocytes and in unselected cells. Our results are of pivotal importance for the future application of amniotic fluid derived stem cells in spina bidfida repair.
脊髓脊膜膨出（MMC）和脊髓裂（MS）被认为是开放性脊柱裂最严重的形式，其特征是在妊娠早期神经管闭合不完全。开放性母胎手术已被确定为修复MMC/MS的一种有前景的替代方法。本研究的主要目标是分离和鉴定源自MMC/MS羊水样本的干细胞。从接受胎儿脊柱裂修复手术的孕妇中获取人羊水样本。我们应用荧光激活细胞分选（FACS）基于cKIT（CD117）表达从羊水细胞异质群体中免疫选择干细胞。然后将cKIT羊水细胞与cKIT和未分选的羊水细胞进行对比，以表征这些细胞的表型。表达cKIT的羊水细胞呈现纺锤形形态，而cKIT阴性的羊水细胞呈圆形。qRT-PCR分析显示，与阴性和未分选的细胞相比，表达cKIT的羊水细胞中KIT和CD90基因的平均mRNA水平显著上调，而CK8基因下调。多能性抗原在分选的cKIT和cKIT羊水细胞以及未分选的细胞中的表达相当。我们的结果对于羊水来源的干细胞在脊柱裂修复中的未来应用至关重要。
