Gan Earn H, Robson Wendy, Murphy Peter, Pickard Robert, Pearce Simon, Oldershaw Rachel
Institute of Genetic MedicineNewcastle University, International Centre for Life, Central Parkway, Newcastle upon Tyne, UK
Endocrine UnitRoyal Victoria Infirmary, Newcastle upon Tyne, UK.
Endocr Connect. 2018 May;7(5):617-629. doi: 10.1530/EC-18-0067. Epub 2018 Apr 5.
The highly plastic nature of adrenal cortex suggests the presence of adrenocortical stem cells (ACSC), but the exact identity of ACSC remains elusive. A few studies have demonstrated the differentiation of adipose or bone marrow-derived mesenchymal stem cells (MSC) into steroid-producing cells. We therefore investigated the isolation of multipotent MSC from human adrenal cortex.
Human adrenals were obtained as discarded surgical material. Single-cell suspensions from human adrenal cortex ( = 3) were cultured onto either complete growth medium (CM) or MSC growth promotion medium (MGPM) in hypoxic condition. Following expansion, their multilineage differentiation capacity was evaluated. Phenotype markers were analysed by immunocytochemistry and flow cytometry for cell-surface antigens associated with bone marrow MSCs and adrenocortical-specific phenotype. Expression of mRNAs for pluripotency markers was assessed by q-PCR.
The formation of colony-forming unit fibroblasts comprising adherent cells with fibroblast-like morphology were observed from the monolayer cell culture, in both CM and MGPM. Cells derived from MGPM revealed differentiation towards osteogenic and adipogenic cell lineages. These cells expressed cell-surface MSC markers (CD44, CD90, CD105 and CD166) but did not express the haematopoietic, lymphocytic or HLA-DR markers. Flow cytometry demonstrated significantly higher expression of GLI1 in cell population harvested from MGPM, which were highly proliferative. They also exhibited increased expression of the pluripotency markers.
Our study demonstrates that human adrenal cortex harbours a mesenchymal stem cell-like population. Understanding the cell biology of adrenal cortex- derived MSCs will inform regenerative medicine approaches in autoimmune Addison's disease.
肾上腺皮质具有高度可塑性,提示存在肾上腺皮质干细胞(ACSC),但其确切身份仍不明确。一些研究已证明脂肪或骨髓来源的间充质干细胞(MSC)可分化为类固醇生成细胞。因此,我们研究了从人肾上腺皮质中分离多能间充质干细胞的方法。
人肾上腺作为废弃手术材料获取。将来自人肾上腺皮质(n = 3)的单细胞悬液在低氧条件下接种于完全生长培养基(CM)或间充质干细胞生长促进培养基(MGPM)中进行培养。扩增后,评估其多向分化能力。通过免疫细胞化学和流式细胞术分析表型标志物,检测与骨髓间充质干细胞和肾上腺皮质特异性表型相关的细胞表面抗原。通过q-PCR评估多能性标志物的mRNA表达。
在CM和MGPM的单层细胞培养中均观察到形成了由具有成纤维细胞样形态的贴壁细胞组成的集落形成单位成纤维细胞。源自MGPM的细胞显示出向成骨和成脂细胞谱系的分化。这些细胞表达细胞表面间充质干细胞标志物(CD44、CD90、CD105和CD166),但不表达造血、淋巴细胞或HLA-DR标志物。流式细胞术显示,从MGPM收获的细胞群体中GLI1表达显著更高,且这些细胞具有高度增殖性。它们还表现出多能性标志物表达增加。
我们的研究表明,人肾上腺皮质中存在类似间充质干细胞的细胞群体。了解肾上腺皮质来源的间充质干细胞的细胞生物学将为自身免疫性艾迪生病的再生医学方法提供信息。