Department of Obstetrics and Gynecology, the Third Affiliated Hospital, Soochow University;
Department of Obstetrics and Gynecology, Nanjing First Hospital, Nanjing Medical University.
J Vis Exp. 2022 Aug 23(186). doi: 10.3791/64065.
Umbilical cord mesenchymal stem cells (UC-MSCs) are an important cell source for regenerative medicine. UC-MSCs can be isolated from the umbilical cord Wharton's jelly, as well as from the umbilical arteries and umbilical vein. They are known as perivascular stem cells obtained from umbilical arteries (UCA-PSCs), perivascular stem cells obtained from the umbilical vein (UCV-PSCs), and mesenchymal stem cells obtained from Wharton's jelly (WJ-MSCs). UCA-PSCs and UCV-PSCs are pericytes derived from perivascular regions that are progenitors of MSCs. Isolation and culture of the three kinds of cells is an important source for studying stem cell transplantation and repair. The present protocol focuses on the isolation and culture of cells through mechanical separation, adherent culture, and cell crawling out. Through this technique, the three different types of stem cells can be derived. Cell surface markers were detected by flow cytometry. The stem cells were detected for multilineage differentiation potential by adipogenic, osteogenic, and neural-like differentiation, which is consistent with the phenotype of MSCs. This experimental protocol expands the source of UC-MSCs. In addition, the cell isolation method provides a basis for further study of regenerative medicine and other applications.
脐带间充质干细胞（UC-MSCs）是再生医学的重要细胞来源。UC-MSCs 可从脐带华通氏胶以及脐动脉和脐静脉中分离得到。它们被称为从脐动脉中获得的血管周干细胞（UCA-PSCs）、从脐静脉中获得的血管周干细胞（UCV-PSCs）和从华通氏胶中获得的间充质干细胞（WJ-MSCs）。UCA-PSCs 和 UCV-PSCs 是源自血管周区域的周细胞，是 MSC 的前体细胞。三种细胞的分离和培养是研究干细胞移植和修复的重要来源。本方案重点介绍通过机械分离、贴壁培养和细胞爬行来分离和培养细胞。通过该技术，可以衍生出三种不同类型的干细胞。通过成脂、成骨和成神经样分化检测细胞表面标志物，检测干细胞的多系分化潜能，这与 MSC 的表型一致。该实验方案扩大了 UC-MSCs 的来源。此外，细胞分离方法为再生医学和其他应用的进一步研究提供了基础。
