Development & Differentiation Research Center, KRIBB, Eoeundong, Yuseong-gu, Daejeon, Republic of Korea.
J Cell Mol Med. 2011 Jan;15(1):152-65. doi: 10.1111/j.1582-4934.2009.00956.x.
Neuropeptide Y (NPY) and NPY receptors are widely expressed in various organs and cell types and have been shown to have pleiotropic functions. However, their presence or role in human embryonic stem cells (hESCs) remains unknown. We now show that undifferentiated hESCs primarily express NPY and its Y1 and Y5 receptors. Inhibition of NPY signalling using either the selective NPY Y1 or Y5 receptor antagonist reduces the maintenance of self-renewal and proliferation of undifferentiated hESCs. We also provide compelling evidence that exogenous NPY supports the long-term growth of undifferentiated hESCs in the absence of feeder cell factors using only knockout serum replacement media. Further, NPY facilitates the use of chemically defined medium made up of N2/B27 supplement and basic fibroblast growth factor (bFGF) for hESC feeder-free culture. Our results indicate that both Y1 and Y5 receptors appear to be involved in the NPY-mediated activation of AKT/protein kinase B and extracellular signal-regulated kinase 1/2 (ERK1/2) in hESCs. Notably, only Y1 receptor, but not Y5 receptor, is responsible for the NPY-induced activation of cAMP-response element binding (CREB) in hESCs. These results provide the first evidence that NPY and its Y1 and Y5 receptors have potential role in maintaining hESC self-renewal and pluripotency. We demonstrate the underlying importance of NPY signalling and its usefulness in the development of a defined and xeno-free culture condition for the large-scale propagation of undifferentiated hESCs.
神经肽 Y（NPY）及其受体广泛表达于多种器官和细胞类型，具有多种生物学功能。然而，它们在人类胚胎干细胞（hESC）中的存在或作用仍不清楚。我们现在发现，未分化的 hESC 主要表达 NPY 及其 Y1 和 Y5 受体。使用选择性 NPY Y1 或 Y5 受体拮抗剂抑制 NPY 信号通路可降低未分化 hESC 的自我更新和增殖维持能力。我们还提供了令人信服的证据，表明外源性 NPY 可在没有饲养细胞因子的情况下，仅使用 knockout 血清替代培养基支持未分化 hESC 的长期生长。此外，NPY 有助于使用由 N2/B27 补充剂和碱性成纤维细胞生长因子（bFGF）组成的化学定义培养基进行 hESC 无饲养细胞培养。我们的结果表明，Y1 和 Y5 受体似乎都参与了 NPY 介导的 hESC 中 AKT/蛋白激酶 B 和细胞外信号调节激酶 1/2（ERK1/2）的激活。值得注意的是，只有 Y1 受体而不是 Y5 受体，负责 NPY 诱导的 hESC 中环磷酸腺苷反应元件结合（CREB）的激活。这些结果首次表明 NPY 及其 Y1 和 Y5 受体在维持 hESC 自我更新和多能性方面具有潜在作用。我们证明了 NPY 信号通路的重要性，以及其在开发无定义和无饲养细胞的培养条件方面的有用性，以实现未分化 hESC 的大规模扩增。
