聚乳酸/羟基磷灰石纳米复合物对MC3T3-E1细胞增殖活性的影响

背景:研究表明聚乳酸/羟基磷灰石复合材料与自然骨结构和性能相似,具有骨传导性和良好的生物相容性。目的:观察聚乳酸/羟基磷灰石纳米复合物与成骨细胞株MC3T3-E1的生物相容性。方法:分别采用普通完全培养基(对照组)与聚乳酸/羟基磷灰石纳米复合物浸提液(实验组)培养第3代成骨细胞株MC3T3-E1,培养3,5,7 d,采用CCK-8法检测细胞的吸光度值;在培养第7,14天检测细胞碱性磷酸酶、骨钙素、Ⅰ型胶原、核心结合因子α1/成骨特异性转录因子表达。将聚乳酸/羟基磷灰石纳米复合物与第3代成骨细胞株MC3T3-E1共培养,培养7,14 d细胞骨架染色及扫描电镜观察细胞在支架材料上的形态。结果与结论:随着时间的延长,成骨细胞株MC3T3-E1的吸光度值明显增加,两组细胞吸光度值比较差异无显著性意义。实验组培养第7天细胞碱性磷酸酶活性高于对照组(P0.05),培养第14天细胞碱性磷酸酶、Ⅰ型胶原、核心结合因子α1/成骨特异性转录因子表达高于对照组(P0.05)。种植7 d后细胞在材料上贴附生长,细胞呈多角形;种植14 d后细胞较7 d前数目明显增多,且完全伸展,呈多角形和梭形。表明聚乳酸/羟基磷灰石纳米复合物对成骨细胞有良好的生物相容性,无细胞毒性。     

新型壳聚糖-胶原支架与牙周膜细胞的生物相容性

背景：目前胶原作为牙周组织工程支架材料仍具有机械强度差、降解速度快等缺点,将其与壳聚糖复合可改善上述问题。 目的：评估新型壳聚糖-胶原支架材料的体外生物相容性。 方法：通过MTT法评估100%,75%,50%,25%壳聚糖-胶原支架材料浸提液对人牙周膜细胞的毒性。选择第4-6代生长状态良好的人牙周膜细胞与壳聚糖-胶原支架共培养,观察细胞在支架上的生长情况,并检测与壳聚糖-胶原支架复合培养前后人牙周膜细胞碱性磷酸酶活性的变化。 结果与结论：新型壳聚糖-胶原支架具有双层结构,一侧表面致密,一侧表面疏松多孔。MTT法检测不同浓度材料浸提液毒性评级为0或1级。扫描电子显微镜及组织学观察可见细胞在壳聚糖-胶原支架上增殖良好,且致密层可起屏障膜作用,阻挡细胞进入支架内部;复合培养24 h后,人牙周膜细胞的碱性磷酸酶活性与复合培养前无明显差异(P > 0.05),复合培养48,72 h后人牙周膜细胞的碱性磷酸酶活性高于复合培养前(P < 0.05)。以上结果提示新型壳聚糖-胶原支架具有良好的生物相容性及屏障功能,可进一步应用于牙周组织工程的研究。     

医用新型Mg-Li-Ca合金材料体外生物相容性及生物活性评价

背景:新型Mg-Li-Ca合金是否具有生物相容性和生物活性,目前还未被证实。目的:通过体外实验评价新型医用Mg-Li-Ca合金的组织相容性及生物活性,初步探讨其作为医用植入材料的可行性。方法:分别采用Mg-Li-Ca合金浸提液、纯Mg浸提液、AZ31B合金浸提液与α-MEM培养液培养第3代小鼠胚胎成骨细胞,培养1,3,5 d,采用MTT法检测细胞A值并计算相对增殖率;培养5,7 d,使用碱性磷酸酶试剂盒检测细胞碱性磷酸酶活性。将第3代小鼠胚胎成骨细胞与Mg-Li-Ca合金、纯Mg、AZ31B合金分别共培养于24孔板,培养1 d后扫描电镜观察材料表面黏附及增殖情况。结果与结论:Mg-Li-Ca合金、纯Mg和AZ31合金对成骨细胞的细胞毒性为Ⅰ级,无细胞毒性,且Mg-Li-Ca合金对细胞毒性明显小于纯Mg和AZ31B合金,对成骨细胞生长增殖无显著影响,呈现出良好的生物相容性。Mg-Li-Ca合金和纯Mg对成骨细胞正常合成碱性磷酸酶无影响,具有良好的生物活性;AZ31B对成骨细胞正常合成碱性磷酸酶有显著影响。成骨细胞可在Mg-Li-Ca合金、纯Mg和AZ31合金表面正常黏附生长。表明新型Mg-Li-Ca合金有望成为新型骨科植入材料。     

聚乳酸-壳聚糖纤维/羟基磷灰石-硅酸钙复合支架材料的细胞相容性

背景:实践证明,有机和无机材料单独应用都不是理想的支架材料。聚乳酸具有良好的生物相容性、生物降解性和生物吸收性,聚乳酸类复合材料将成为21世纪最重要的生物复合材料之一。目的:观察复合支架材料聚乳酸-壳聚糖纤维/羟基磷灰石-硅酸钙对成骨细胞黏附、增殖、分化的影响。方法:取新生24h内Wistar大鼠的颅盖骨,采用改良胶原酶消化法进行成骨细胞原代培养。通过倒置相差显微镜、苏木精-伊红染色、碱性磷酸酶染色、钙结节茜素红染色对获得的细胞进行生物学特性的观察与鉴定。然后将第3代细胞与聚乳酸/壳聚糖纤维、聚乳酸-壳聚糖纤维/硅酸钙、聚乳酸-壳聚糖纤维/羟基磷灰石-硅酸钙三种支架材料体外复合培养。培养3,6,9d后,采用倒置相差显微镜观察材料周边的细胞形态,通过碱性磷酸酶活性测定法和MTT法观察3种支架材料对细胞分化、增殖的影响。结果与结论:三种材料均有利于成骨细胞的黏附、生长、分化、增殖,而聚乳酸-壳聚糖纤维/羟基磷灰石-硅酸钙复合支架材料较聚乳酸/壳聚糖纤维、聚乳酸-壳聚糖纤维/硅酸钙支架材料促进成骨细胞的分化增殖效果更好,证实其生物相容性好,有望成为一种新型的骨组织工程支架材料。     

硼硅酸盐对成骨细胞体外生物活性的影响

背景：硼硅酸盐不仅可通过矿化作用形成羟基碳酸盐磷灰石层,而且具有强化学反应活性,可促进骨细胞再生。 目的：通过体外培养实验观察硼硅酸盐生物玻璃对兔成骨细胞生长行为的影响。 方法：根据 ISO10993-12：2007 的要求制备硼硅酸盐生物玻璃初次浸提液与二次浸提液。分离培养兔骨髓间充质干细胞,取第2代细胞诱导生成成骨细胞。取第5-15代成骨细胞,分别以硼硅酸盐生物玻璃初次浸提液、硼硅酸盐生物玻璃二次浸提液与α-MEM培养基培养,观察硼硅酸盐生物活性玻璃对成骨细胞增殖、蛋白合成、碱性磷酸酶活性、细胞凋亡及细胞横向与纵向迁移的影响。 结果与结论：初次浸提液组与二次浸提液组成骨细胞增殖优于α-MEM培养基组(P < 0.05),且初次浸提液组成骨细胞增殖优于二次浸提液组(P < 0.05)。初次浸提液组成骨细胞总蛋白含量高于二次浸提液组与α-MEM培养基组(P < 0.05)。3组间成骨细胞碱性磷酸酶活性、凋亡率、横向迁移距离及Transwell 中穿膜细胞数比较差异均无显著性意义。表明硼硅酸盐生物玻璃具有良好的细胞相容性,对成骨细胞增殖有一定的良性调节作用。  中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

新型有序纳米介孔生物活性玻璃体外复合培养成骨细胞生物相容性研究

目的评价新型有序纳米介孔生物活性玻璃(80SMBG)对成骨细胞的生物相容性。方法以Novabone为对照,用大鼠成骨细胞与80SMBG浸提培养液、Novabone浸提培养液和正常的空白培养液培养,分别于3、4、5d用四唑盐(MTT)比色法计数计算细胞相对增殖度,用6级毒性分类法评级,同时用偶氮偶联法(PNPP)染色,分析碱性磷酸酶活性的变化。结果80SMBG毒性评级为0～1级,对成骨细胞分化无明显的抑制作用。结论80SMBG的体外生物活性较高,对体液微环境的影响轻微,细胞相容性好,作为骨组织修复替代材料或骨组织工程支架材料具有较高的研究和应用价值。     

纳米钛酸钙对鼠成骨细胞增殖与分化的影响

背景：以不同修饰剂制备的纳米钛酸钙具有良好的骨传导性、生物相容性及生物活性。 目的：观察纳米钛酸钙材料对成骨细胞增殖、分化的影响。 方法：分别以聚乙二醇、十六烷基三甲基溴化铵、柠檬酸钠为修饰剂制备纳米钛酸钙,同时制备无修饰剂的纳米钛酸钙。将制备的4组纳米钛酸钙分别配制为0.1,1.0,10 g/L的浸提液,MTT方法检测材料浸提液对乳鼠成骨细胞增殖及碱性磷酸酶活性的影响,以正常培养的细胞为对照。 结果与结论：以聚乙二醇、十六烷基三甲基溴化铵、柠檬酸钠为修饰剂制备的纳米钛酸钙,无修饰剂的纳米钛酸钙在1.0,10 g/L质量浓度下可明显促进成骨细胞的增殖;上述4组纳米钛酸钙浸提液在1.0,10 g/L质量浓度下,培养3,6,9 d的细胞A值也高于对照组(P < 0.05,P < 0.01),培养6,9 d的细胞碱性磷酸酶活性值均高于对照组(P < 0.05,P < 0.01)。表明以聚乙二醇、十六烷基三甲基溴化铵、柠檬酸钠、无修饰剂制备的钛酸钙对成骨细胞无毒性作用,可促进成骨细胞的增殖和分化。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程全文链接：     

掺锶聚磷酸钙的细胞相容性

背景:研究显示,以可降解生物陶瓷聚磷酸钙为基材,加入微量锶元素,通过调控材料结构如孔径、孔隙率等性能,可制备出有足够力学强度并可控降解的掺锶聚磷酸钙。目的:观察成骨细胞与掺锶聚磷酸钙的细胞相容性。方法:将兔成骨细胞与不同含锶质量分数(0～100%)的掺锶聚磷酸钙体外复合培养,进行形态学和功能测定。MTT法检测掺锶聚磷酸钙中成骨细胞的活力,并测定成骨细胞碱性磷酸酶活性。结果与结论:在倒置显微镜、扫描电子显微镜和激光共聚焦显微镜下观察到成骨细胞在掺锶聚磷酸钙的表面和孔隙内大量黏附、增殖和生长,并且分泌细胞外基质及细胞表面的大量微绒毛,显示成骨细胞在掺锶聚磷酸钙材料上生长形态良好;同时通过MTT和碱性磷酸酶测定表明细胞功能良好,其中含锶1%掺锶聚磷酸钙细胞增殖活性及碱性磷酸酶活性最高,对成骨细胞无毒性,具有良好的细胞相容性。     

聚醚醚酮-羟基磷灰石-碳纤维复合材料的生物相容性研究

目的对聚醚醚酮(PEEK)-羟基磷灰石(HA)-碳纤维(CF)复合材料的生物相容性进行评价。方法应用注塑法制备不同质量分数的PEEK-HA-CF复合材料,选择70PEEK-20HA-10CF复合材料(PEEK、HA、CF质量分数分别为70%、20%、10%)为实验组,75PEEK-05HA-20CF复合材料(PEEK、HA、CF质量分数分别为75%、5%、20%)为阳性对照组,70PEEK-30CF(PEEK、CF质量分数分别为70%、30%)为阴性对照组。从新生SD大鼠颅盖骨提取原代成骨细胞,培养至第2代,用于实验。测定不同质量分数的PEEK-HA-CF复合材料的黏附性、增殖能力、细胞形态及相关碱性磷酸酶(ALP)、Ⅰ型胶原(COL-Ⅰ)、骨钙素(OCN)mRNA含量,评价复合材料的生物相容性。结果荧光显微镜下CF和HA均匀分布在PEEK中。与对照组相比,各组细胞黏附性差异无统计学意义(P 0.05);实验组第5天、第7天光密度(OD)分别为1.88±0.08、2.83±0.15,阳性对照组第5天、第7天OD值为1.55±0.09、2.56±0.12,阴性对照组第3天、第5天OD值分别为1.47±0.03、2.46±0.06;实验组增殖力明显高于对照组(P 0.05)。ALP mRNA、COL-ⅠmRNA含量在第14天时达到高峰;在第5、7天时,实验组高于对照组(P 0.05),而两对照组间差异无统计学意义(P 0.05);第14天时,实验组ALP mRNA、COL-ⅠmRNA含量高于对照组(P 0.05),且阳性对照组高于阴性对照组(P 0.05),而3组间OCN mRNA表达差异无统计学意义(P 0.05)。扫描电子显微镜观察发现,细胞在复合材料上生长良好,并随着时间的延长,细胞在材料表面达到融合,形成矿化颗粒。结论 PEEK-HA-CF复合材料具有良好的生物相容性和生物活性。     

人骨形成蛋白2基因成骨诱导作用研究

目的制备一种具有成骨活性的生物材料。方法将人BMP2-pcDNA3．1质粒添加到壳聚糖-胶原海绵支架材料中,然后将大鼠成骨细胞和成纤维细胞分别接种在该复合材料中,体外培养细胞-支架材料复合物,另外将该材料植入BALBc小鼠皮下,以添加了pcDNA3．1质粒的壳聚糖-胶原海绵支架材料为对照。10 d以后,取出细胞-支架材料复合物和皮下植入物,分别检测样品的碱性磷酸酶活性和钙含量。结果与对照相比发现,体外培养细胞-支架材料复合物中的碱性磷酸酶活性和钙含量均显著提高;小鼠皮下植入物的碱性磷酸酶活性和钙含量也分别提高了61．5％和16．2％。结论添加了人骨形成蛋白2基因的壳聚糖-胶原海绵支架材料具有成骨诱导活性,这种材料为骨组织工程中的成骨诱导可能提供一种新的方法。     

Ad-LIF-OSM双基因转染饲养细胞对脐血造血细胞体外增殖和分化的影响

目的 构建表达人白血病抑制因子(LIF)和抑瘤素M(OSM)双基因的WI-38人胚肺成纤维细胞,并以此细胞作为饲养层细胞观察其对CD34+.造血干/祖细胞体外增殖和分化的影响.方法 以双启动子转移载体pAdTrack-CMV-LIF-polyA+promoterΔ为基础,将OSM基因片段酶切后插入,构建出转移质粒pAdTrack-CMV-LIF-polyA+promoterΔ -OSM.将构建正确的转移质粒与腺病毒骨架质粒共转化,获得重组腺病毒载体pAdEasy-1-pAdTrack-CMV-LIF-polyA+promoter△-OSM,通过包装,收获重组腺病毒(AdLIF-OSM).将重组腺病毒感染饲养层细胞,经RT-PCR、ELISA法检测外源基因在细胞中的表达;体外与CD34+造血干/祖细胞共培养后,通过Transwell法和细胞计数检测,比较各实验组干/祖细胞体外扩增与分化情况.结果 测序结果显示重组载体中的LIF和OSM基因序列正确;转基因饲养层细胞中能检测到外源LIF和OSM基因的转录和表达;外源LIF和OSM基因在造血干/祖细胞体外培养中能够发挥作用.结论 成功构建携带人LIF和OSM的双基因重组腺病毒载体(Ad-LIF-OSM),Ad-LIF-OSM在造血干/祖细胞体外培养的过程中能够有效地扩增CD34+造血干/祖细胞,并延缓其分化.     

Human umbilical cord blood cells express neurotrophic factors

Freshly isolated or culture-expanded human umbilical cord blood mononuclear cells (CBMNCs) have been known to express neural phenotypes in vitro and to differentiate into neural cells and improve neurological function recovery after being administrated into rodent models of neurological diseases. However, the mechanism of action remains unclear. The present study observed that CBMNCs expressed higher level mRNAs of several neurotrophic factors than adult peripheral blood mononuclear cells (PBMCs). In addition, a significantly increase in the levels of brain-derived neurotrophic factor (BDNF) and neurotrophin-4/5 (NT4/5) was found in culture supernatants of CBMNCs compared to that of PBMNCs. These findings indicate that CBMNCs express several neurotrophic factors and suggest that the neurotrophic factors secreted by CBMNCs may be responsible for amelioration of central nervous system deficits in animal models after CBMNC administration.     

Upregulation of LMP1 Expression by Histone Deacetylase Inhibitors in an EBV Carrying NPC Cell Line

OBJECTIVES: In about 60% of Epstein-Barr virus (EBV) carrying nasopharyngeal carcinomas (NPC) LMP1 expressing cells can be detected. The frequency of LMP1 positive cells and the expression level varies from cell to cell in the different tumors. Cell lines derived from EBV positive NPCs loose the virus during in vitro culture. The in vitro infected NPC cell line TWO3-EBV used in our study carries the neomycin-resistance gene containing EBV and expresses low level of LMP1. With this cell line it was thus possible to study the regulation of LMP1 expression by modification of chromatin acetylation state. STUDY DESIGN: The TWO-EBV cell line was treated with n -butyrate (NB) or trichostatin A (TSA). RESULTS: Shown by immunoblotting, the LMP1 level was elevated in the treated samples. Already 2 h after TSA exposure LMP1 expression was higher and it increased up to 24 h. Immunofluorescence staining showed that nearly all cells were LMP1 positive. Neither EBNA2 nor BZLF1 were induced. Tested first 2 h after the treatment, acetylated histone H3 and H4 were already detectable, and their level increased up to 8 h. Chromatin immunoprecipitation (ChIP) verified that the LMP1-promoter (LMP1p) (ED-L1) was acetylated after TSA treatment. CONCLUSION: EBV carrying epithelial cells do not express EBNA-2. We showed that LMP1 expression was upregulated by histone deacetylase inhibitors in an in vitro infected, EBV carrier NPC cell line.     

In vitro apoptosis effects of GnRHII on endometrial stromal cells from patients with endometriosis

Purpose: To study the effect of gonadotropin-releasing hormone II (GnRHII) on the cell apoptosis of ectopic, eutopic and normal endometrial stromal cells cultured in vitro from endometriosis patients, and to provide theoretical basis for exploring new treatments for endometriosis (EMs). Methods: Ectopic, eutopic and normal endometrial stromal cells were isolated, cultured and identified in vitro, then treated with different concentrations of GnRHII (0, 10^-10 M, 10^-8 M and 10^-6 M). Cell apoptosis was detected by Hoechst staining and flow cytometry. Results: GnRHII increased apoptosis in ectopic, eutopic and normal stromal cells in a dosage-dependent manner (P<0.05), and apoptosis of ectopic stroma cells was significantly higher than that of eutopic and normal cells (P<0.05); apoptosis in eutopic and normal cells had no different (P>0.05). Conclusion: GnRHII can significantly induce apoptosis in ectopic, eutopic and normal endometrial stromal cells from patients with endometriosis, especially to the ectopic.     

Suppression by Human Placental Protein 14 of Natural Killer Cell Activity

ABSTRACT: Human decidua of early pregnancy contains considerable numbers of CD3^? CD56⁺ natural killer (NK) cells. In this study, two major protein products of the decidua, placental protein 14 (PP14) and placental protein 12 (PP12), were tested for the ability to regulate human NK cell activity. In vitro overnight exposure to PP14 of blood lymphocytes or purified large granular lymphocytes (LGL) resulted in suppression of cytotoxicity against K562 target cells in a 4-h ⁵¹Cr release assay. The NK inhibition was dependent on concentrations of PP14, being detectable at 5 μg/ml and reaching maximum at 50 μg/ml. Manifestation of PP14-induced NK suppression required 18-h contact with NK cells. The suppression of NK activity by PP14 was not abolished by indomethacin. In a target binding assay the number of PP14-treated LGL binding to K562 was comparable to that of untreated ones. By contrast with PP14, PP12 produced no effects on NK cells. These results indicate that PP14 suppresses the function of NK cells, which might be involved in prevention of maternal immune rejection of fetus at the fetomaternal interface.     

Critical contribution of CD80 and CD86 to induction of anterior chamber-associated immune deviation

Intraocular inoculation of antigens induces anterior chamber-associated immune deviation (ACAID), which is mediated by development of regulatory T cells in response to antigen-presenting cells (APC) pre-conditioned by intraocular transforming growth factor-beta (TGF-beta). In this study, we examined the involvement of T-cell co-stimulatory molecules in this process. To mimic the intraocular APC, thioglycollate-elicited peritoneal exudate cells (PEC) were pre-treated with TGF-beta in vitro. Expression of CD80, CD86, OX40 ligand (OX40L) and CD70 was analyzed by flow cytometry. Contribution of these molecules to co-stimulatory activity of TGF-beta-treated PEC on antigen-stimulated T-cell proliferation and cytokine production was determined by inhibition with blocking antibodies in vitro. Contribution of CD80 and CD86 to induction of ACAID was determined by the administration of blocking antibodies at intraocular antigen inoculation in vivo. TGF-beta-treated PEC expressed CD80 and CD86 but not OX40L or CD70. Antigen-stimulated T cells proliferated and produced IL-10, but not IFN-gamma, in response to co-stimulation by TGF-beta-treated PEC, which was abrogated by blocking antibodies against CD80 and CD86. Induction of regulatory cells mediating ACAID was abolished by in vivo blockade of CD80 and CD86. The present results indicated that CD80 and CD86 play a critical role in induction of ACAID, possibly by co-stimulating expansion and IL-10 production of regulatory T cells in response to TGF-beta-conditioned APC.     

HSV-TK/GCV自杀基因系统治疗宫颈癌的研究

目的探讨HSV-TK/GCV自杀基因系统对人宫颈癌细胞系Hela体外及体内杀伤作用及其产生的旁观者效应.方法采用脂质体转染法将GINaTK载体转入包装细胞PA317.取病毒上清液感染人宫颈癌细胞Hela,得到带有HSV-TK基因的Hela/TK细胞,并将其分别用于体外和体内实验.结果载体HSV-TK导入了PA317细胞.体外实验结果显示,当Hela/TK细胞数占混合细胞10%时,低浓度(10μg/ml)的GCV就可将50%左右的肿瘤细胞杀死.体内实验结果显示GCV可明显抑制Hela/TK细胞在BALB/C小鼠体内的肿瘤形成.经GCV治疗后,肿瘤体积分别较对照组肿瘤体积缩小约11.1%、30.6%和47.2%(均P＜0.001);RT-PCR检测HSV-TK基因在肿瘤组织中有表达;实验组肿瘤组织与对照组相比存在明显的病理学改变.结论逆转录病毒可介导HSV-TK基因转入人宫颈癌细胞Hela并获稳定表达,HSV-TK/GCV自杀基因系统在体内外对宫颈癌细胞均有杀伤作用,且存在明显的旁观者效应.     

Human ovarian tumour-derived chaperone-rich cell lysate (CRCL) elicits T cell responses in vitro

Tumour-derived chaperone-rich cell lysate (CRCL), which is made up of numerous heat shock proteins, has been used successfully to generate tumour-specific T cell responses and protective immunity against a wide range of murine tumours. In this study, we have investigated the potency of human ovarian cancer-derived CRCL to activate dendritic cells (DC) and to generate tumour-specific T cells in vitro. CRCL was generated from primary ovarian cancers and SKOV3-A2, a HER2/neu, Wilm's tumour gene 1 (WT1) and human leucocyte antigen (HLA)-A2 positive human ovarian tumour cell line. Peripheral blood mononuclear cells from both HLA-A2(+) healthy donors and HLA-A2(+) ovarian cancer patients were stimulated weekly with autologous DC loaded with ovarian tumour-derived CRCL. After four to six stimulations in vitro, specific cytokine secretion and cytotoxicity were measured. CRCL promoted interleukin (IL)-12 secretion and enhanced the immunostimulatory capacity of DC. T cells from healthy controls and from ovarian cancer patients secreted higher amounts of interferon-gamma following in vitro restimulation with ovarian cancer-derived CRCL than with HER2/neu or WT1 peptide-pulsed DC. We were also able to generate cytotoxic T lymphocyte activity against cancer-specific antigens such as HER2/neu and WT1 from all healthy donors, but from only one of the four ovarian cancer patients with bulky disease. These preliminary results substantiate further the concept that CRCL may prove to be a potent adjuvant for women suffering from ovarian cancer and that this personalized vaccine may be a promising approach for active immunotherapy.