犬内皮与平滑肌细胞的短期静态联合培养

目的:为获得组织工程化自体血管,观察体外静态培养条件下犬内皮细胞与平滑肌细胞联合培养组织学及形态学的特征。方法:实验于2004-07/2005-06在首都医科大学宣武医院外科院级实验室完成。①实验材料:雄性杂种犬,3个月龄,体质量8～12kg。②实验方法:贴块法及酶解法对犬内皮细胞及平滑肌细胞进行原代分离培养及扩增,将第Ⅱ代平滑肌细胞以1×109L-1的密度种植于胶原膜上培养13d,再将第Ⅱ代内皮细胞接种于生长平滑肌细胞的胶原膜上2d。③实验评估:行苏木精-伊红染色同时扫描电镜和透射电镜观察平滑肌细胞和内皮细胞在胶原载体上联合培养后的形态。结果:苏木精-伊红染色见平滑肌细胞较均匀的分布于支架材料表面及内部;扫描电镜下,平滑肌细胞可以在胶原载体材料上生长,增殖明显并在短期内形成多层细胞。内皮细胞与平滑肌细胞联合培养2d就可在平滑肌细胞层表面获得连续的单层内皮细胞层。结论:在短期静态培养条件下犬的血管平滑肌细胞和内皮细胞可以在胶原载体材料上形成具有两层细胞结构的组织工程化动脉血管组织片。     

具有周向微槽结构管状血管支架制备及诱导平滑肌细胞的取向生长

背景:对小口径血管组织工程化而言,平滑肌细胞的周向排列要求彻底改变以前支架的简单多孔结构,代之以能够诱导血管平滑肌细胞三维周向取向和排列新型微观结构。目的:观察微槽结构对平滑肌细胞体外定向诱导的影响。方法:用静电纺丝、熔融纺丝并利用溶剂/非溶剂和热压的方式制得了具有两层管壁、外壁具有周向微沟槽结构的仿生管状血管支架,用胶原蛋白固定改性后,在其上种植人脐静脉血管平滑肌细胞。扫描电镜和荧光显微镜观察支架不同缠绕角度对平滑肌细胞定向诱导能力的影响。结果与结论:①选择比例为5∶95的氯仿/乙醇溶液,浸润时间为5s,可以使乳酸-羟基乙酸共聚物电纺纤维和乳酸-ε-己内酯共聚物熔纺纤维之间形成很好的粘连,形成支架。②通过碱降解使支架表面含有羧基,以1-(二甲基胺丙基)-3-乙基碳化二亚胺为缩合剂在支架表面接枝胶原。X射线光电子能谱证实了支架表面胶原大分子的存在。③当纤维之间的编织角度为30°即网孔尺寸适当时,细胞能在支架内部和表面大面积生长。④具有两层管壁结构的仿生管状血管支架具有良好的细胞相容性,其表面周向微槽结构对平滑肌细胞的取向排列具有明显的诱导作用。提示在电纺层外面再熔纺缠绕降解聚合物是制备管状仿生血管支架的可行方法。血管平滑肌细胞能沿着纤维暨微沟槽方向一致取向排列。     

人脐带内皮细胞与平滑肌细胞的分离培养特照

目的：如何便捷有效地获取种子细胞是构建组织工程皮肤、角膜、血管等成功与否的关键，实验对人胎儿脐带内皮细胞和平滑肌细胞的分离、纯化、培养扩增技术进行探讨。方法：实验于2006—11在暨南大学医学院完成。①实验材料：剖宫产正常胎儿脐带由暨南大学第一临床学院提供，产妇均知情同意。②实验方法：配制培养液，A液为在M199培养液中加入20％胎牛血清、20mg，L肝素、10μg／L碱性成纤维细胞生长因子，B液为在M199培养基中加入10％胎牛血清、250mg／L G418。取脐带15-20cm，采用胶原酶“灌注消化法”获取脐带内皮细胞制成悬液，静置0．5h后利用成纤维细胞短时间内贴壁的特点，把尚未贴壁细胞吸出重新接种于含有A液的培养瓶，初步去除部分成纤维细胞，细胞贴壁24h后将培养液换为B液，继续培养3d，去除成纤维细胞，然后再用A液扩大培养。从消化完脐静脉内皮细胞的脐带中剥取脐动脉，刮除内皮细胞，将血管条剪成小块均匀贴于培养瓶底，培养瓶内加入含20％小牛血清的DMEM培养液2mL，缓慢竖直培养瓶，以培养液刚未浸没培养块为最佳，放入37℃、体积分数为0．05的CO2培养箱内干涸8h后，将培养瓶轻轻翻转，使植块刚浸入培养液中，绝对静置3d，以后每3d换液1次，待植块周围生长晕的细胞融合成片时，即可传代，传代后DMEM培养液的血清浓度由原来的20％改为5％，且在培养液中加入0．2L肝素。⑧实验评估：通过倒置相差显微镜观察细胞生长情况，分别以第Ⅷ因子、α-肌动蛋白免疫组织化学染色法鉴定内皮细胞和平滑肌细胞。结果：①内皮细胞的生长观察：接种24h后，镜下可见刚贴壁的细胞呈圆形，逐渐转变为多角形或梭形，胞间可见有成纤维细胞混杂，加入B液3d后成纤维细胞逐渐死亡。传代细胞呈“铺路石”样，细胞周围特别是近胞核处可见明显光晕，为典型的内皮细胞生长特点。②内皮细胞第Ⅷ因子相关抗原的检测：胞浆丰富且有棕黄色颗粒，细胞核不着色，呈阳性反应。⑧平滑肌细胞的生长观察：镜下组织块贴壁后5～7d可见有少数细胞从植块边缘游出，第10-15天植块周围形成明显的细胞生长晕．细胞呈长梭形、带状或三角状，有1-4个细胞核，可见半透明颗粒，第20-25天细胞密集、平行排列，形成“峰谷样”结构，是平滑肌细胞的特征性生长表现。④平滑肌细胞特异性α-肌动蛋白单克隆抗体检测：胞浆内可见棕黄色细丝，为特异性α-平滑肌肌动蛋白，细胞核不着色。结论：以胎儿脐带作为种子细胞的来源，成功分离后通过调节培养液的成分，既能排除成纤维细胞的污染又可以令内皮细胞和平滑肌细胞快速增殖。     

制备可缓释血管内皮生长因子的聚乳酸-聚乙醇酸支架及其与血管平滑肌细胞的生物相容性

目的:制备一种新型的可缓释血管内皮生长因子的可降解聚乳酸-聚乙醇酸支架,探讨其与血管平滑肌细胞的细胞相容性。方法:实验于2006-12/2007-05在解放军第四军医大学西京医院心内科实验室(省部级重点实验室)完成。以明胶微球作为缓释载体,加载血管内皮生长因子和聚乳酸-聚乙醇酸复合构建组织工程血管支架并考察体外释放效果。采用体外培养的兔血管平滑肌细胞种植在可缓释血管内皮生长因子的可降解聚乳酸-聚乙醇酸支架膜片上,用相差显微镜观察细胞的生长情况,绘制细胞生长曲线,并用MTT法测定细胞增殖情况。实验过程中对动物处置符合动物伦理学要求。结果:血管内皮生长因子在体外释放14d以上,血管平滑肌细胞在可缓释血管内皮生长因子的可降解聚乳酸-聚乙醇酸支架膜片上生长良好;MTT法检测细胞增殖情况显示,可缓释血管内皮生长因子的可降解聚乳酸-聚乙醇酸支架组平滑肌细胞增殖较快,细胞较为活跃(P<0.05)。结论:可缓释血管内皮生长因子的可降解聚乳酸-聚乙醇酸支架制备方法简便易行,生长因子活性得以保存,对血管平滑肌细胞具有良好的相容性,可以用于组织工程血管的进一步构建。     

Defining smooth muscle cells and smooth muscle injury

For 3 decades, terms such as synthetic phenotype and contractile phenotype have been used to imply the existence of a specific mechanism for smooth muscle cell (SMC) responses to injury. In this issue of the JCI, Hendrix et al. offer a far more precise approach to examining the mechanisms of SMC responses to injury, focused not on general changes in phenotype but on effects of injury on a single promoter element, the CArG [CC(A/T)6GG] box, in a single gene encoding smooth muscle (SM) alpha-actin. Since CArG box structures are present in some, but not all, SMC genes, these data suggest that we may be progressing toward establishing a systematic, molecular classification of both SMC subsets and the response of SMCs to different injuries.     

A biomimetic scaffold for culturing limbal stem cells: a promising alternative for clinical transplantation

Limbal tissues can be cultured on various types of scaffolds to create a sheet of limbal-corneal epithelium for research as well as clinical transplantation. An optically clear, biocompatible, biomimetic scaffold would be an ideal replacement graft for transplanting limbal stem cells. In this study, we evaluated the physical and culture characteristics of the recombinant human cross-linked collagen scaffold (RHC-III scaffold) and compared it with denuded human amniotic membrane (HAM). Optical/mechanical properties and microbial susceptibility were measured for the scaffolds. With the approval of the institutional review board, 2 mm fresh human limbal tissues were cultured on 2.5 x 2.5 cm(2) scaffolds in a medium containing autologous serum in a feeder cell-free submerged system. The cultured cell systems were characterized by morphology and immunohistochemistry for putative stem cells and differentiated cell markers. The refractive index (RI) and tensile strength of the RHC-III scaffold were comparable to human cornea, with delayed in vitro degradation compared to HAM. RHC-III scaffolds were 10-fold less susceptible to microbial growth. Cultures were initiated on day 1, expanded to form a monolayer by day 3 and covered the entire growth surface in 10 days. Stratified epithelium on the scaffolds was visualized by transmission electron microscopy. The cultured cells showed p63 and ABCG2 positivity in the basal layer and were immunoreactive for cytokeratin K3 and K12 in the suprabasal layers. RHC-III scaffold supports and retains the growth and stemness of limbal stem cells, in addition to resembling human cornea; thus, it could be a good replacement scaffold for growing cells for clinical transplantation.     

Effects of 2-deoxy-D-glucose on proliferation of vascular smooth muscle cells and endothelial cells

2-Deoxy-D-glucose (2-DG) is a glucose analogue that has been proposed for cancer therapy due to its cytostatic properties. Its effect on the proliferation of smooth muscle cells and endothelial cells has not been fully clarified. The aims of this study were to investigate the effects of 2-DG on the proliferation of porcine aortic endothelial cells (PAEC) and porcine smooth muscle cells (PSMC), to establish an overview of its dose-dependent inhibitory capacity and to examine whether the short-term incubation of cells with 2-DG has an impact on cell proliferation in culture. Our results showed a dose-dependent significant inhibitory effect on proliferation, which was more pronounced in PSMC than in PAEC. Even after short-term incubation of cells with 2-DG, relevant inhibition of proliferation was documented. The clinical application of 2-DG might be a promising concept by inhibiting cells that show a potentially rapid proliferation in response to non-malignant stimuli, such as smooth muscle cells after intracoronary stenting.     

小鼠主动脉平滑肌细胞及肌源性泡沫细胞与格列本脲共孵育的反应

目的:观察磺酰脲类药物格列本脲对平滑肌细胞及肌源性泡沫细胞内酰基辅酶A:胆固醇酰基转移酶1(ACAT1)酶活性的影响.方法:实验于2003-06/2005--07在华中科技大学同济医学院实验中心完成.①用组织贴块法培养小鼠主动脉平滑肌细胞,采用两步超速离心法制备低密度脂蛋白后,运用CuCl2氧化法制备氧化低密度脂蛋白,采用100 mg/L的氧化低密度脂蛋白作用于平滑肌细胞72 h.建立泡沫细胞模型.②在平滑肌细胞和泡沫细胞分别加入格列本脲,使终浓度为200μmol/L,孵育24 h,用放射性同位素标记法检测ACAT1酶活性.结果: ①平滑肌细胞转变为泡沫细胞后,ACAT1酶活性上升(P<0.05).②平滑肌细胞被格列本脲干预后,ACAT1酶活性无明显变化(P>0.05);肌源性泡沫细胞加入格列本脲后,ACAT1酶活性下降(P<0.05)结论:格列本脲能抑制肌源性泡沫细胞内ACAT1的酶活性,抑制平滑肌细胞向泡沫细胞的转化,有可能成为防止动脉粥样硬化的有效药物.     

体表海绵状静脉畸形中内皮细胞及平滑肌细胞分布与表型对血管塑形的影响

目的:研究内皮细胞、平滑肌细胞在体表海绵状静脉畸形中的分布与表型,并分析其发病机制和研究结果对疾病治疗和功能康复的意义。方法:实验于1996-06/2000-08在第二军医大学整形外科实验室完成。畸形组织25根,正常中小静脉各12根。苏木精-伊红染色观察比较畸形血窦壁和中小静脉壁中内皮细胞、平滑肌细胞的分布并计数分析。Envision法免疫组化染色,观察CD31,α平滑肌肌动蛋白在畸形和中小静脉中的分布。结果:畸形血窦壁中平滑肌细胞数量少,排列紊乱,平滑肌细胞与内皮细胞的比例显著低于微小静脉壁。畸形中CD31表达与中小静脉相似,但α平滑肌肌动蛋白的表达明显低于中小静脉,排列紊乱。结论:海绵状静脉畸形血窦壁中内皮细胞和平滑肌细胞发育不均衡,平滑肌细胞表型异常,导致血窦壁薄弱而在血窦内血液的压力下不断扩张,引起血管塑形障碍,这可能是病变发生和进展的原因。闭塞病理性血窦、阻断病变进展过程从而尽可能保留功能是临床治疗的重点。     

人多发性骨髓瘤细胞株RPMI8226细胞对共培养内皮细胞的作用

目的 在体外共培养体系中研究人多发性骨髓瘤(MM)细胞对人正常内皮细胞的作用.方法 建立人MM细胞株RPMI8226细胞与正常人脐静脉内皮细胞(HUVEC)的体外共培养体系;以同期单独培养的HUVEC为对照,用电子显微镜和光学显微镜对与RPMI8226细胞共培养的HUVEC进行细胞超微结构和细胞形态学观察;用刮痕迁移实验、管状结构形成实验评价体外内皮细胞血管新生的能力;并以Western blot和流式细胞术分别检测脑源性神经营养因子(BDNF)、Endoglin与TrkB、Tie2、β3,整合素、血管细胞黏附分子-1蛋白的表达.结果 与同期单独培养的正常内皮细胞对照相比,共培养后的部分内皮细胞胞体伸展并首尾排列成一行;超微结构出现胞核增大、核仁增大、核质比例增大,内质网疏松、变形,细胞表面纤毛减少;共培养体系中细胞管状结构形成数量和迁移细胞数较单独培养体系分别增加了112%和136%;BDNF、Endoglin、TrkB、Tie2、β3整合素和血管细胞黏附分子-1蛋白的表达亦明显上调.结论 MM细胞与人正常内皮细胞共培养后,内皮细胞有明显的趋瘤特征和较强的血管新生能力,推测MM患者骨髓中骨髓瘤细胞通过对相邻内皮细胞的作用促进血管新生,且新生血管的内皮细胞性质与行为不同于正常内皮细胞.     

与人脐动脉平滑肌细胞共培养的人脐静脉内皮细胞的生物学特性

目的:通过共培养人脐静脉内皮细胞(human umbilical venous endothelial ceils,HUVEC)与人脐动脉平滑肌细胞(hunlan umbilical arterial smooth muscle cells,HUASMC),初步探讨观察共培养的HUVEC的形态和增殖特性及经皮冠状动脉成形术(percutaneous transluminal coronary angioplasty,PTCA)后再狭窄的机制。 方法:用共培养装置将体外同时培养在一个培养孔中的HUVEC和HUASMC分离。在倒置显微镜下观察单独培养和共培养HUVEC的生长状态和形态;采用免疫荧光法标记细胞,流式细胞仪测定荧光强度和细胞周期。 结果:共培养的HUVEC逐渐转变为长梭形;单独培养的HUVEC仍保持鹅卵石样。单独培养的和共培养的HUVEC处于细胞周期GO/G1期和G_2+s期的百分率为:(70±3)％和(81±5)％:(22±4)％和(14± 4)％。二者均有Cyclin D的表达,但后者的表达显著低于前者。结论:共培养的HUVEC低血清干预48h形态逐渐变为长梭形,更接近于在体形状;而单独培养的HUVEC仍保持原鹅卵石状。此外,前者的增殖活性也显著低于后者。     

RNA干扰内皮细胞分泌白细胞介素6影响平滑肌细胞的迁移

背景:血管支架置入后靶血管部位易发生炎症反应。目的:利用siRNA技术抑制内皮细胞白细胞介素6的生成,观察其对平滑肌细胞迁移的影响。方法:采用RT-PCR测定脂多糖刺激EA.HY926细胞表达白细胞介素6 mRNA的时间梯度与浓度梯度,针对白细胞介素6构建短发卡状siRNA真核表达载体pGensil-1.1-白细胞介素6,通过lipofectamine 2000转染EA.HY926,抑制其白细胞介素6的产生。结果与结论:pGensil-1.1-白细胞介素6转染EA.HY926细胞后,脂多糖刺激下EA.HY926细胞表达的白细胞介素6 mRNA及蛋白明显减少。共培养模型中,转染pGensil-1.1-白细胞介素6的EA.HY926细胞作用下,人脐静脉平滑肌细胞表达的基质金属蛋白酶9 mRNA及蛋白明显降低,结晶紫染色显示人脐静脉平滑肌细胞迁移数量减少。说明siRNA技术可抑制内皮细胞白细胞介素6的生成,并通过降低平滑肌细胞基质金属蛋白酶9的表达减弱平滑肌细胞的迁移能力。     

RNA干扰内皮细胞分泌白细胞介素6影响平滑肌细胞的迁移

背景：血管支架置入后靶血管部位易发生炎症反应。目的：利用siRNA技术抑制内皮细胞白细胞介素6的生成,观察其对平滑肌细胞迁移的影响。方法：采用RT-PCR测定脂多糖刺激EA.HY926细胞表达白细胞介素6 mRNA的时间梯度与浓度梯度,针对白细胞介素6构建短发卡状siRNA真核表达载体pGensil-1.1-白细胞介素6,通过lipofectamine 2000转染EA.HY926,抑制其白细胞介素6的产生。结果与结论：pGensil-1.1-白细胞介素6转染EA.HY926细胞后,脂多糖刺激下EA.HY926细胞表达的白细胞介素6 mRNA及蛋白明显减少。共培养模型中,转染pGensil-1.1-白细胞介素6的EA.HY926细胞作用下,人脐静脉平滑肌细胞表达的基质金属蛋白酶9 mRNA及蛋白明显降低,结晶紫染色显示人脐静脉平滑肌细胞迁移数量减少。说明siRNA技术可抑制内皮细胞白细胞介素6的生成,并通过降低平滑肌细胞基质金属蛋白酶9的表达减弱平滑肌细胞的迁移能力。     

IQGAP1-dependent scaffold suppresses RhoA and inhibits airway smooth muscle contraction

The intracellular scaffold protein IQGAP1 supports protein complexes in conjunction with numerous binding partners involved in multiple cellular processes. Here, we determined that IQGAP1 modulates airway smooth muscle contractility. Compared with WT controls, at baseline as well as after immune sensitization and challenge, Iqgap1^–/– mice had higher airway responsiveness. Tracheal rings from Iqgap1^–/– mice generated greater agonist-induced contractile force, even after removal of the epithelium. RhoA, a regulator of airway smooth muscle contractility, was activated in airway smooth muscle lysates from Iqgap1^–/– mice. Likewise, knockdown of IQGAP1 in primary human airway smooth muscle cells increased RhoA activity. Immunoprecipitation studies indicated that IQGAP1 binds to both RhoA and p190A-RhoGAP, a GTPase-activating protein that normally inhibits RhoA activation. Proximity ligation assays in primary airway human smooth muscle cells and mouse tracheal sections revealed colocalization of p190A-RhoGAP and RhoA; however, these proteins did not colocalize in IQGAP1 knockdown cells or in Iqgap1^–/– trachea. Compared with healthy controls, human subjects with asthma had decreased IQGAP1 expression in airway biopsies. Together, these data demonstrate that IQGAP1 acts as a scaffold that colocalizes p190A-RhoGAP and RhoA, inactivating RhoA and suppressing airway smooth muscle contraction. Furthermore, our results suggest that IQGAP1 has the potential to modulate airway contraction severity in acute asthma.     

内皮前体细胞与平滑肌细胞联合培养构建组织工程血管:最适比例验证

目的:由内皮前体细胞分化的内皮细胞与成熟平滑肌细胞联合培养可以为组织工程血管的形成提供更接近天然的条件.实验拟进一步证明两种细胞联合培养可促进血管内皮细胞生长,以及两种细胞的最适比例.方法:实验于2006-01/2007-05在复旦大学上海医学院分子生物学实验室完成.①实验材料:新鲜脐带取自国际第一妇婴保健医院,产妇知情同意.②实验方法及评估:从人脐动脉中用组织块培养法分离并原代培养血管平滑肌细胞,免疫荧光方法鉴定其平滑肌肌动蛋白表达情况;采用密度梯度离心法分离脐血中的内皮前体细胞,经体外定向诱导分化和免疫荧光方法鉴定其表型;无菌条件下制作Ⅰ型胶原凝胶,在其上以3:1～4:1的比例混匀联合培养内皮前体细胞与平滑肌细胞,并观察两种细胞联合培养时的形态,分析两种细胞最合适的种植比例;免疫荧光方法观察在Ⅰ型胶原凝胶联合培养中CD31、vWF的表达以及内皮细胞的生长情况.结果:①原代培养的平滑肌细胞呈典型"波峰谷"形态,荧光染色平滑肌肌动蛋白呈阳性.②脐血来源的内皮前体细胞定向诱导分化为内皮细胞后,呈现出"铺路石"样形态,表达CD31、vWF,结合荆豆凝集素,提示具备成熟内皮细胞特性.③在Ⅰ型胶原凝胶上两种细胞增殖力均旺盛,与平滑肌细胞以3:1～4:1的比例种植在I型胶原凝胶培养一段时间后,内皮前体细胞可从平滑肌细胞处得到支持,形成血管样网络结构.④共培养1周后,CD31与vWF阳性细胞即内皮前体细胞分化而来的内皮细胞互相连接,形成环形结构.结论:内皮前体细胞和平滑肌细胞以3:1～4:1共培养的模式可以促进微血管样结构的形成.     

Basic fibrobrast growth factor induces the secretion of vascular endothelial growth factor by human aortic smooth muscle cells but not by endothelial cells

BACKGROUND: Endothelial cell dysfunction and smooth muscle cell (SMC) proliferation are major events in atherogenesis. Both cells are a source of growth factors that mediate cellular proliferation and chemotaxis. Inappropriate production of, and/or response to, these growth factors (such as vascular endothelial growth factor, VEGF, and basic fibroblast growth factor (bFGF)) may contribute to atherogenesis and therefore to disease progression. METHODS: Production of VEGF and its soluble receptor (sFlt-1) by human SMCs and human umbilical endothelial cells (HUVECs) after stimulation with bFGF were examined by ELISA of cell culture media and by Western blotting. RESULTS: Smooth muscle cells produced significantly more VEGF than HUVECs (P<0.05) after 24 h of culture with bFGF levels > or =0.001 microg mL(-1). bFGF induced dose-dependent production of VEGF by SMCs, where maximum production was present in 1 microg mL(-1) of bFGF. Conversely, the SMCs produced less sFlt-1 than HUVECs (P<0.05). However, bFGF induced dose-dependent phosphorylation of Flt1 and another VEGF receptor, KDR, in HUVECs but not SMCs. There was no VEGF or sFLT-1 after 6 h of culture in any dose of bFGF in either type of cell. CONCLUSIONS: Differences in the production of VEGF and sFlt-1 by SMCs and HUVECs are consistent with the role of these cells in angiogenesis. Induction of VEGF production and expression by bFGF in these cells indicates that this growth factor may participate in angiogenesis indirectly by the induction of VEGF. The production of sFlt-1 by both cell types is in agreement with the notion that sFlt-1 may be involved in the regulation of VEGF activity. Additionally, the ability of bFGF to induce dose-dependent phosphorylation of KDR in HUVECs highlights the important role of bFGF in VEGF-mediated angiogenic processes.     

与人脐静脉内皮细胞共培养的人脐动脉平滑肌细胞生物学特性研究

目的:通过共培养人脐静脉内皮细胞(humanumbilicalvenousendothe-lialcells,HUVEC)与人脐动脉平滑肌细胞(humanumbilicalarterialsmoothmusclecells,HUASMC),初步探讨共培养的HUASMC的形态和增殖特性。方法:用共培养装置将体外同时培养在一个培养孔中的HUVEC和HUASMC分离。在倒置显微镜下观察单独培养和共培养HUASMC的生长状态和形态;采用免疫荧光法标记细胞,流式细胞仪测定荧光强度和细胞周期。结果:共培养的HUASMC由长梭状变肥大,处于静止期状态;单独培养的HUASMC仍保持长梭形。共培养的和单独培养的HUASMC处于细胞周期G2+S期和G0/G1期的百分率为:(85±4)%和(69±3)%;(11±2)%和(20±3)%。两者均有CyclinD的表达,但后者的表达显著高于前者。结论:单独培养的HUASMC无血清干预48h后仍保持长梭形,而共培养的HUASMC逐渐变肥大,进入静止期形态。此外,前者的增殖活性也显著高于后者。