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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 observe the biological characteristics of human corpus cavernosum smooth muscle cells (hCCMCs) in three-dimensional culture system. Methods hCCMCs were isolated in vitro and purified by differential adhesion method. Immunohistochemistry was used to detect the expression of α-smooth muscle actin (α-SMA), and the hCCMCs were identified. On the basis of monolayer adherent culture, hCCMCs were cultured in collagen gel to form a three-dimensional culture system, and the morphological structure and proliferation of hCCMCs were studied. Results The cells of primary cultured hCCMCs were heterogeneous. After being purified by differential adherence, the cells were homogenized. Immunohistochemistry showed that most of the cells were positive for α-SMA. In the three-dimensional culture system, hCCMCs grow three-dimensionally at different levels. The number of hCCMCs cultured in three-dimensional culture system for 10 days showed no significant proliferation (P> 0.05), which was significantly lower than that in adherent culture (P <0.01). Conclusion The hCCMCs three-dimensional culture system can observe the growth and proliferation status of hCCMCs under three-dimensional conditions and study the interaction between hCCMCs and microenvironment. It is expected that a simpler and more controllable in vivo experimental alternative will be found for related research.