兔垂体干细胞的培养鉴定及分化

Culture,identification,and differentiation of rabbit pituitary stem cells

背景:脑垂体是人体重要的内分泌器官。多种疾病可引起脑垂体损伤,如脑垂体瘤、垂体功能减退症等。垂体干细胞因具有自我更新和多向分化潜力的特性,有望成为一种新的治疗方式修复受损的垂体。目的:采用悬浮细胞球培养法分离培养垂体干细胞,鉴定其增殖及分化能力。方法:采用悬浮细胞球培养法从新生新西兰白兔垂体中分离培养垂体干细胞,观察其形态特征;免疫荧光细胞染色检测垂体干细胞标志蛋白SOX2及Nestin的表达;采用EdU标记法检测垂体干细胞增殖能力;经贴壁及诱导分化后,通过ELISA法检测培养基中的激素水平。结果与结论:通过悬浮细胞球培养法可成功分离得到垂体干细胞球,且具有较强的增殖能力;干细胞特异性标记物SOX2和Nestin阳性表达;经诱导分化后培养基中促肾上腺皮质激素、促甲状腺激素、促生长激素、促黄体生成激素、促卵泡素、催乳素水平显著升高(P<0.001),具有较强的分化能力。

BACKGROUND:The pituitary gland is an important endocrine organ in the body.Certain diseases can cause damage to the pituitary gland,such as pituitary adenoma and abnormal hormone secretion.Pituitary stem cells,due to their self-renewal and multi-directional differentiation potential,are expected to become a new therapeutic approach for repairing damaged pituitary glands.OBJECTIVE:To isolate and culture pituitary stem cells using the suspension cell ball culture method and identify their proliferation and differentiation ability.METHODS:Pituitary stem cells were isolated and cultured from the pituitary gland of newborn New Zealand white rabbits using the suspension cell ball culture method,and their morphological characteristics were observed.Immunofluorescence cytochemistry was used to detect the expression of pituitary stem cell markers SOX2 and Nestin.EdU labeling method was utilized to detect the proliferative ability of pituitary stem cells.After adherent and induced differentiation,the hormone levels in the culture medium were detected by ELISA.RESULTS AND CONCLUSION:Pituitary stem cell spheres could be successfully isolated by the suspension cell ball culture method,with strong proliferative ability.Positive expression of stem cell-specific markers SOX2 and Nestin was found in the cultured cells.After induction and differentiation,adrenocorticotropic hormone,thyroid hormone,growth hormone,luteinizing hormone,follicle-stimulating hormone,and prolactin levels significantly increased in the medium(P<0.001),with strong differentiation ability.