摘要
目的观察人阴茎海绵体平滑肌细胞(hCCMCs)在三维培养系统中的生物学特性。方法体外分离hCCMCs,并用差速贴壁法进行纯化。免疫组化的方法检测α-平滑肌肌动蛋白(α-SMA)的表达,进行hCCMCs的鉴定。在单层贴壁培养的基础上,将hCCMCs在胶原凝胶中培养,形成三维培养系统,并对hCCMCs的形态结构、增殖情况进行研究。结果原代培养的hCCMCs细胞形态不均一,经过差速贴壁法纯化后,细胞形态均一,免疫组化显示大多数细胞α-SMA阳性表达。三维培养系统中,hCCMCs在不同层面上呈三维生长。hCCMCs在三维培养系统中培养10d后,细胞数量无明显增殖(P>0.05),明显低于贴壁培养条件下细胞增殖速率(P<0.01)。结论 hCCMCs三维培养系统可以观察hCCMCs在三维条件下的生长、增殖状况,研究hCCMCs与微环境的相互作用关系,有望为相关研究找到一种更为简单、可控性更强的体内实验替代方法。
Objective To analyze the biological properties of human corpus cavernosum smooth muscle cells （hCCMCs） under three-dimensional culture system. Methods hCCMCs were isolated in vitro, and purified by gradient velocity sedimentation, hCCMCs were identified by the expression of α-smooth muscle actin （ α-SMA）. On the basis of adherent monolayer culture, hCCMCs were seeded into collagen gel for three-dimensional culture Morphology and proliferation of hCCMCs were studied under 3D state. Results The morphology of the primary hCCMCs was heterogeneous. After gradient velocity sedimentation, the morphology of hCCMCs became homogeneous. Positive expression of α-SMA was detected in most of hCCMCs by immunohistochemistry, hCCMCs grew at different levels in three-dimensional culture system.No significant proliferation was found in hCCMCs after 10 days three-dimensional culture（P〉0.05）, Proliferation rate of them was significantly lower than that of adherent monolayer cells（P〈0.01）. Conclusion Three-dimensional culture system could he used to detect the growth and proliferation of hCCMCs under the three-dimensional conditions, and study the interaction of hCCMCs and micro-environment. This research was expected to find a simpler and more controllable alternative to related experiments in vivo.
出处
《中国男科学杂志》
CAS
CSCD
2010年第2期12-15,共4页
Chinese Journal of Andrology
关键词
阴茎
肌细胞
平滑肌
细胞培养技术
人
penis
myocytes, smooth muscle
cell culture techniques
human
