CAJ | 학술논문

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>O.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.
목적 관찰인음경해면체평활기세포(hCCMCs)재삼유배양계통중적생물학특성.방법 체외분리hCCMCs,병용차속첩벽법진행순화.면역조화적방법검측α-평활기기동단백(α-SMA)적표체,진행hCCMCs적감정.재단층첩벽배양적기출상,장hCCMCs재효원응효중배양,형성삼유배양계통,병대hCCMCs적형태결구、증식정황진행연구.결과 원대배양적hCCMCs세포형태불균일,경과차속첩벽법순화후,세포형태균일,면역조화현시대다수세포α-SMA양성표체.삼유배양계통중,hCCMCs재불동층면상정삼유생장.hCCMCs재삼유배양계통중배양10d후,세포수량무명현증식(P>O.05),명현저우첩벽배양조건하세포증식속솔(P<0.01).결론 hCCMCs삼유배양계통가이관찰hCCMCs재삼유조건하적생장、증식상황,연구hCCMCs여미배경적상호작용관계,유망위상관연구조도일충경위간단、가공성경강적체내실험체대방법.
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 be 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.