Department of Surgery, Division of Endocrine Surgery, University of Michigan Health System, Ann Arbor, MI 48109-5654, USA.
Surgery. 2010 Dec;148(6):1186-9; discussion 1189-90. doi: 10.1016/j.surg.2010.09.021.
Hypoparathyroidism is the most frequent permanent complication of thyroid surgery. Our hypothesis is that human precursor cells in culture can be differentiated into parathyroid cells and used to reconstitute function. Human embryonic stem cells (hESCs) are a stable model to study differentiation into parathyroid-like cells. In prior work, the BG01-hESC line was stimulated to form parathyroid-like cells. This cell line is no longer available, however, and additional studies were needed to confirm and extend prior observations.
Increasing concentrations of fetal bovine serum and timed exposure to Activin A were used to differentiate H1-hESC into parathyroid-like cells. The potential benefit of Sonic hedgehog exposure on parathyroid-like cell development also was evaluated by serial alterations of culture conditions. Calcium-sensing receptor (CaSR), GCM2, and PTH expression (RT-PCR) and PTH protein secretion (ELISA) were used as markers of differentiated cells.
We successfully modified our prior protocol to generate cells that express CaSR, GCM2, and PTH RNA from undifferentiated H1-hESC. The cells also secreted PTH.
We replicated parathyroid differentiation using H1-hESC cells. Our data advance the project toward in vitro differentiation of precursor cells isolated from individual patients for autotransplantation.
甲状旁腺功能减退症是甲状腺手术后最常见的永久性并发症。我们的假设是，培养中的人前体细胞可以分化为甲状旁腺细胞，并用于重建功能。人类胚胎干细胞（hESC）是研究分化为甲状旁腺样细胞的稳定模型。在之前的工作中，BG01-hESC 系被刺激形成甲状旁腺样细胞。然而，该细胞系已不再可用，需要进行额外的研究来证实和扩展之前的观察结果。
使用胎牛血清浓度递增和激活素 A 定时暴露来将 H1-hESC 分化为甲状旁腺样细胞。还通过连续改变培养条件来评估 Sonic hedgehog 暴露对甲状旁腺样细胞发育的潜在益处。钙敏感受体 (CaSR)、GCM2 和 PTH 表达（RT-PCR）和 PTH 蛋白分泌（ELISA）被用作分化细胞的标志物。
我们成功修改了之前的方案，从未分化的 H1-hESC 中生成表达 CaSR、GCM2 和 PTH RNA 的细胞。这些细胞还分泌 PTH。
我们使用 H1-hESC 细胞复制了甲状旁腺的分化。我们的数据推进了该项目，即从个体患者中分离出前体细胞进行体外分化，然后进行自体移植。
