Abstract

With CRISPR/Cas9 gene editing technology, human umbilical cord mesenchymal stem cells (HUMSCs) were directionally induced to differentiate into parathyroid cells. The study provides the theoretical and experimental basis for the application of HUMSCs in the clinical treatment of parathyroid hypofunction. In this study the CRISPR/Cas9 gene editing technology was used to introduce the PTH gene into HUMSCs to induce the differentiation of HUMSCs. After 28 days of induction culture, HUMSCs edited by CRISPR/Cas9 showed morphological changes. PTH expression was detected by quantitative real-time PCR (qRT-PCR), western blotting and immunohistochemistry, the expression level of PTH in parathyroid adenoid cells obtained after CRISPR/Cas9 gene editing was significantly increased. The expression levels of CASR, GCM2 and PTH were 98.9%, 93.6% and 97%, respectively. This experiment verified that editing the PTH gene with the CRISPR/Cas9 system could directionally induce human umbilical cord mesenchymal stem cells to differentiate into parathyroid cells.

摘要

利用CRISPR/Cas9基因编辑技术，定向诱导人脐带间充质干细胞（HUMSCs）分化为甲状旁腺细胞。该研究为HUMSCs在临床治疗甲状旁腺功能减退症中的应用提供了理论和实验依据。本研究采用CRISPR/Cas9基因编辑技术将PTH基因导入HUMSCs，诱导HUMSCs分化。诱导培养28天后，CRISPR/Cas9编辑的HUMSCs出现形态变化。通过定量实时PCR（qRT-PCR）、蛋白质印迹和免疫组织化学检测PTH的表达，CRISPR/Cas9基因编辑后获得的甲状旁腺腺样细胞中PTH的表达水平显着增加。CASR、GCM2和PTH的表达水平分别为98.9%、93.6%和97%，分别。本实验验证了用CRISPR/Cas9系统编辑PTH基因可以定向诱导人脐带间充质干细胞分化为甲状旁腺细胞。