The differentiation of amniotic fluid stem cells into sweat glandlike cells is enhanced by the presence of Sonic hedgehog in the conditioned medium
The differentiation of amniotic fluid stem cells into sweat glandlike cells is enhanced by the presence of Sonic hedgehog in the conditioned medium
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条件培养基中存在 Sonic Hedgehog 可增强羊水干细胞向汗腺样细胞的分化
DOI：
10.1111/exd.13062
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发表时间：
2016-09-01
影响因子：
3.6
通讯作者：
Qin, Mingde
中科院分区：
文献类型：
--
作者：
Liang, Hansi;Sun, Qing;Qin, Mingde
在患者遭受严重的全层烧伤后，目前的治疗方法不能导致汗腺结构和功能的完全自我再生。因此，寻找新的方法获得足够的功能性汗腺细胞以恢复皮肤功能是很重要的。在这项研究中，我们利用人汗腺(HSG)细胞的条件培养液(CM)诱导CD117+人羊水干细胞(HAFS)分化为汗腺样细胞(HAFS-SG)。实时定量聚合酶链式反应和免疫荧光染色证实汗腺相关基因外胚抗原-A(EDA)、外营养不良蛋白-A受体(EDAR)、角蛋白8(K8)和癌胚抗原(CEA)的表达。透射电子显微镜分析表明，HSG细胞的典型细胞结构--微绒毛也可见于Hafs-SG细胞膜上。我们对乙酰胆碱(Ach)的钙反应的测试证明，Hafs-SG细胞具有以类似于正常汗腺的方式对Ach作出反应的潜力。三维培养是刺激Hafs-SG细胞形成管状结构，驱动Hafs-SG细胞向更高阶段成熟的有效途径。我们还发现，表皮生长因子提高了分化的效率，而声波刺猬是影响汗腺分化的CM的重要因素。我们的研究为进一步研究诱导干细胞分化为汗腺样细胞的新方法提供了基础。
After patients suffer severe full-thickness burn injuries, the current treatments cannot lead to the complete self-regeneration of the sweat gland structure and function. Therefore, it is important to identify new methods for acquiring sufficient functional sweat gland cells to restore skin function. In this study, we induced CD117+ human amniotic fluid stem (hAFS) cells to differentiate into sweat glandlike (hAFS-SG) cells based on the use of conditioned medium (CM) from the human sweat gland (hSG) cells. Real-time PCR and immunofluorescent staining were used to confirm the expression of the sweat gland-related genes Ectodysplasin-A (EDA), Ectodysplasin-A receptor (EDAR), keratin 8 (K8) and carcino-embryonic antigen (CEA). Transmission electron microscopy analysis shows that microvilli, the cellular structures that are typical for hSG cells, can also be observed on the membrane of the hAFS-SG cells. Our test for the calcium response to acetylcholine (Ach) proved that hAFS-SG cells have the potential to respond to Ach in a manner similar to normal sweat glands. A three-dimensional culture is an effective approach that stimulates the hAFS-SG cells to form tubular structures and drives hAFS-SG cells to mature into higher stage. We also found that epidermal growth factor enhances the efficiency of differentiation and that Sonic hedgehog is an important factor of the CM that influences sweat gland differentiation. Our study provides the basis for further investigations into novel methods of inducing stem cells to differentiate into sweat glandlike cells.