摘要
目的研究使用共培养系统将骨髓间充质干细胞分化为有分泌功能的颌下腺腺泡细胞的可行性。方法分离成年SD大鼠骨髓间充质干细胞于体外培养、纯化和鉴定,同时分离新生SD大鼠的颌下腺细胞于体外培养、纯化和鉴定,取颌下腺腺泡细胞第二代和间充质干细胞第一代置于共培养系统诱导干细胞定向分化。使用免疫细胞化学法分析骨髓间充质干细胞是否表达淀粉酶抗体。结果纯化后的颌下腺腺泡细胞呈铺路石样形态,原代至第5代培养基及细胞内均可检测到淀粉酶表达;体外培养的骨髓间充质干细胞呈梭形,呈束状、漩涡状排列,可成功诱导成脂肪细胞,油红-O染色显示含有脂滴;共培养后的骨髓间充质干细胞表达淀粉酶抗体,约30%的骨髓间充质干细胞在共培养1周后转化为腺泡细胞,约50%的骨髓间充质干细胞在共培养2周后转化为腺泡细胞。结论成功构建大鼠颌下腺腺泡细胞和骨髓间充质干细胞的体外培养体系,并证明大鼠骨髓间充质干细胞具有转化为腺泡细胞的能力,因此骨髓间充质干细胞将有望应用于涎腺组织工程。
Objective To investigate the feasibility of transdifferentiation of bone marrow parenchymal stem cells（MSCs）into functional submandibular gland acinar cells （SMGCs） using a co-culture systerm. Method Bone marrow parenchyrnal stem cells were isolated from adult SD rats and cultured, purified, identified in vitro, at the same time, submandibular gland aeinar cells were isolated from neonatal rats and cultured, purifide, identified in vitro, then MSCs of passage 1 were co-cultured with SMGCs of passage 2, to induce the MSCs to transdifferentiate into acinar cells. Since the acinar cell was the only cell which had the function of secreting amylase, the transdifferentiation of bone marrow cell was identified by immunocytochemical analysis with amylase. Result Cobblestone-like submandibular gland acinar cells were found after purification, and amylase was successfully detected both in culture medium and cells of passage 0-5 respectively. The MSCs were in fusiform shape, arranged in bundles or whorls, and can successfully be transdifferentiated into adipocytes, which was confirmed by using Oil Red-O staining. Expression of amylase, which may indicate successful transdifferntiation of MSCs into acinar cells, was found in 30% of MSCs after being co-cultured for 1 week, and in 50% after co-culturing for 2 and 3 weeks. Conclusion It has been successful to culture SMGCs and MSCs in vitro, and it has also been demonstrated that the the rat MSCs have the potentiality of being transdifferentiated into acinar cells, therefore it is applicable in salivary gland tissue engineering.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2009年第5期562-565,共4页
Medical Journal of Chinese People's Liberation Army
基金
广东省自然科学基金博士启动项目(06300692)
关键词
细胞分化
间质干细胞
颌下腺
组织工程
cell differentiation
mesenchymal stem cell
submandibular gland
tissue engineering
