不同培养液对体外培养的小鼠胸腺上皮细胞增殖的影响

分离纯化小鼠胸腺上皮细胞（mTEC）,以DMEM、DMEM/F12、RPMI 1640 3种基础培养液加10%胎牛血清完全培养液进行培养, MTT法和流式细胞术分别检测mTEC的增殖及细胞周期,筛选出最适合mTEC生长的培养液。结果表明,DMEM培养的mTEC增殖快,细胞增殖指数明显高于DMEM/F12、RPMI 1640培养液,表明DMEM是mTEC体外培养的最佳培养液。     

小鼠卵巢颗粒细胞的优化培养

为了找到小鼠卵巢颗粒细胞更加合适的体外生长环境,本研究分别向分离的小鼠卵巢颗粒细胞中加入无血清RMPI1640、含10%血清RMPI1640、无血清DMEM（高糖）、含10%血清DMEM（高糖）及含15%血清DMEM（高糖）的培养液对小鼠卵巢颗粒细胞进行体外培养。试验经观察结果显示,DMEM（高糖）总体培养效果好于RMPI1640,且含15%血清的DMEM（高糖）是较理想的培养用液。     

小鼠腹腔巨噬细胞的分离培养与鉴定

目的建立一种小鼠腹腔巨噬细胞分离培养的简便方法。方法以无血清的DMEM培养液灌洗小鼠腹腔,分离获取小鼠腹腔巨噬细胞,在含有10%成牛血清的DMEM培养液中培养。采用倒置显微镜观察细胞形态,台盼蓝染色计算存活率,瑞氏染色计算纯度。结果获得高纯度的巨噬细胞,具备巨噬细胞的形态特征。结论该方法是一种简单可行的分离巨噬细胞的方法。     

不同型牛血清对家兔原核胚序贯培养体外发育的影响

采用不同型血清分别添加到TCM199和mTCM199培养液、RPMI1640和mRPMI1640培养液中，对兔原核期受精卵进行了体外序贯培养，并对各组间不同时期发育率进行了分析比较。结果显示：体外培养至第72h时，3个体外序贯培养体系间8-细胞胚率、桑椹胚率差异不显著（P〉0．05），当体外培养至囊胚时，100mL／L NBS＋TCM199（mTCM199）培养体系、100mL／L NBS＋RPMI1640（mRPMI1640）培养体系的囊胚率均显著低于100mL／L FBS＋RPMI1640（mRPMI1640）培养体系的囊胚率（三组的囊胚率依次是27．3％、35．9%、97．2%，P〈0．01）。但前两组之间差异不显著。结果表明，不同型血清对兔早期胚胎体外正常发育具有很大影响，序贯培养中添加国产NBS的培养液的培养效果明显低于添加进口FBS的培养液的培养效果。     

小鼠腹腔巨噬细胞的分离培养与鉴定

目的建立一种小鼠腹腔巨噬细胞分离培养的简便方法.方法以无血清的DMEM培养液灌洗小鼠腹腔,分离获取小鼠腹腔巨噬细胞,在含有10％成牛血清的DMEM培养液中培养.采用倒置显微镜观察细胞形态,台盼蓝染色计算存活率,瑞氏染色计算纯度.结果获得高纯度的巨噬细胞,具备巨噬细胞的形态特征.结论该方法是一种简单可行的分离巨噬细胞的方法.     

小鼠腹腔巨噬细胞的分离培养与鉴定

目的:建立一种小鼠腹腔巨噬细胞分离培养的简便方法。方法:以无血清的DMEM培养液灌洗小鼠腹腔,分离获取小鼠腹腔巨噬细胞,在含有10%成牛血清的DMEM培养液中培养。采用倒置显微镜观察细胞形态,台盼蓝染色计算存活率,瑞氏染色计算纯度。结果:获得高纯度的巨噬细胞,具备巨噬细胞的形态特征。结论:该方法是一种简单可行的分离巨噬细胞的方法。     

弓形体RH株在3种传代细胞中增殖情况的比较

为建立有效适用的弓形体体外培养方法,本试验利用Vero、PK15、RK13 3种传代细胞和DMEM、MEM、RPM I1640三种培养液分别对RH株弓形体速殖子进行培养,并比较了虫体的增殖和活力情况。结果显示,利用Vero细胞DMEM培养液培养弓形体速殖子,虫体增殖快、活力强,是理想的培养方法。     

淫羊藿苷对体外金黄色葡萄球菌感染小鼠巨噬细胞和淋巴细胞的免疫调节效应

为分析淫羊藿苷(ICA)对体外金黄色葡萄球菌感染小鼠巨噬细胞和淋巴细胞的免疫调节效应。制备腹腔巨噬细胞和脾淋巴细胞悬液,用含10%胎牛血清的RPMI-1640(培养基)培养,在培养液中分别加入不同浓度(终浓度为0、1.0、1.8、3.0、4.2和4.8μmol/L)的ICA。经不同时间培养后,感染金黄色葡萄球菌,再培养不同时间,MTT法检测巨噬细胞的吞噬和淋巴细胞的增殖,ELISA法检测淋巴细胞白细胞介素-2和白细胞介素-10的分泌,流式细胞仪检测淋巴细胞周期。结果表明:ICA能够提高金黄色葡萄球菌感染的巨噬细胞的吞噬能力,促进淋巴细胞的增殖,提高淋巴细胞进入S期的百分率,促进淋巴细胞白细胞介素-2和白细胞介素-10的分泌。结果显示:ICA对金黄色葡萄球菌感染具有免疫调节效应。     

不同培养基对小鼠黄体细胞体外培养效果的影响

对未性成熟的雌性小鼠注射孕马血清促性腺激素和人绒毛膜促性腺激素,在适当的时间摘取小鼠卵巢进行卵巢黄体细胞的体外培养,培养用液为含10%血清的DMEM和含10%血清的RPMI 1640。结果发现,用前者培养的细胞在形态、规则程度和数量上都优于后者,为小鼠黄体细胞体外培养液的选择和进一步研究提供了参考。     

不同培养方法对鸡精原干细胞体外培养效果的影响

本研究从孵化至16 d的鸡胚睾丸中分离获取精原干细胞（Spermatogonial stem cells,SSCs）,比较3种培养液、2种处理（有无饲养层）对SSCs生长的影响,并对SSCs保持特性进行鉴定。结果表明：在无饲养层细胞存在的条件下,DMEM、TCM199和RPMI1640培养液中,SSCs的存活时间分别为6.5、6.0和3.5 d,DMEM和TCM199培养液之间差异不显著（P〉0.05）,但两者与RPMI1640之间均存在极显著差异（P〈0.01）。在有饲养层细胞存在的条件下,在DMEM、TCM199和RPMI1640培养液中,SSCs的存活时间分别为45.5、38.0和14.0 d,三者相互之间存在着极显著的差异（P〈0.01）。在6种培养体系中,SSCs在培养体系Ⅳ中的存活时间和AKP阳性克隆率分别为（45.5±3.20）d和0.31±0.46,极显著地高于其他5种培养体系（P〈0.01）。SSCs在培养体系Ⅳ中传代至1、2和3代时,AKP阳性克隆率分别为31.6%、20%和18%。SSCs形成的克隆,经AKP活性检测和SSEA-1免疫染色,均呈阳性。传代至第3代的SSCs,其染色体组型保持不变,为2n=78。这些结果表明,SSCs在以DMEM为基础培养基的培养体系Ⅳ中培养至第3代时,仍保持干细胞未分化特性。     

多房棘球绦虫-泡球蚴的体外培养研究

按照多房棘球绦虫幼虫-泡球蚴培养的培养基（RPMI-1640、M199和MEM）分为3组：Ⅰ组为含10％胎牛血清的RPMI-1640;Ⅱ组为含10％胎牛血清的MEM;HI组为含10％胎牛血清的M199。将泡球蚴在3种细胞培养液中进行培养,观察其存活、生长以及发育情况。结果显示,培养9d的泡球蚴的成活率分别为：Ⅰ组90．10％、Ⅱ组50．25％、Ⅲ组22．03％;成囊率分别为：Ⅰ组57．12％、Ⅱ组63．15％、Ⅲ组48．17％;头节外翻率分别为：Ⅰ组98．28％、Ⅱ组88．65％、Ⅲ组75．50％。可见,大多数虫体在早期向囊发育,一部分虫体头节外翻,并伴有规律的伸缩运动,但随时间的延长虫体运动减缓,又向囊蚴发育。通过对多房棘球绦虫泡球蚴的体外培养,初步表明合有10％小牛血清的细胞培养基RPMI-1640较适合泡球蚴的生长发育,为研究寄生虫发育提供了最基本的数据资料。     

Preliminary observations on thein-vitro culture of metacestodes ofTaenia saginata

Metacestodes ofTaenia saginata were cultured in a diphasic medium consisting of a disrupted solid phase of coagulated calf serum and a fluid phase of hepes buffered RPMI-1640 enriched with sodium pyruvate and foetal calf serum. The growing tapeworms formed segments, which showed early development of sexual organs. This culture technique gave better results than methods using monophasic media or an intact solid phase when assessed in terms both of the survival of the cestodes and of five parameters measured at the end of the culture period.     

In vitro cultivation of a newly recognized Babesia sp. in dogs in North Carolina

A novel large Babesia sp. from an infected dog was cultivated in vitro by microaerophilous stationary phase culture methodology. A primary culture initiated in enriched RPMI-1640 medium supplemented with 40% canine serum and incubated in a 2% oxygen environment supported parasite growth in vitro. Subsequent subcultures into enriched HL-1 medium with 20% fetal bovine serum also supported parasite propagation. Cultures were successfully introduced to 5% carbon dioxide in air atmosphere at passage 4. To date, the parasites have been continuously cultured through 35 passages, although the parasitemias are low, ranging from 0.2 to 0.3%. Parasites cultured in RPMI with canine serum were cryopreserved and successfully recovered from liquid nitrogen storage. The small subunit ribosomal rRNA gene sequence was identical in blood-derived and culture-derived parasites, differing in a single base position from the previously reported sequence for this Babesia sp. The ultrastructure of the parasite was consistent with that of other large Babesia spp., except that the spherical body contained numerous round particles unlike the inclusions previously described in Babesia spp.     

培养基质中添加酯化葡甘露聚糖和DON对小鼠脾细胞生长和转化的影响

将体外培养的脾细胞分为5组,以RPMI-1640为基础培养基,第1组为空白对照组;第2组为阳性对照组,添加500 μg/L DON;第3组、第4组和第5组在添加500 μg/L DON的基础上分别添加0.05%、0.10%和0.15%的EGM。在培养72 h后,测定各组培养基质中MDA含量、SOD活性、脾细胞存活率和淋巴细胞转化率。结果表明,第1组、第4组和第5组的脾细胞存活率、SOD活性极显著高于第2组(P<0.01);第1组、第4组和第5组的淋巴细胞转化率显著高于第2组(P>0.05);第2组培养液中MDA含量显著高于第1组、第4组和第5组(P<0.05);第1组、第4组和第5组各项指标差异不显著(P>0.05)。由此表明,EGM能很好地吸附DON,减轻DON对小鼠脾细胞的毒性作用;EGM在饲料中的适宜添加剂量为0.10%。     

In vitro platelet release by rat megakaryocytes: effect of heterologous antiplatelet serum

A visual assay to study megakaryocyte platelet release via proplatelet formation in vitro was established. Samples of megakaryocyte-enriched rat bone marrow were incubated (37 C) in RPMI-1640 medium with 15% autologous serum in specially prepared chambers. In the culture system, approximately 6% of megakaryocytes formed proplatelet processes within 24 hours. Inclusion of a heterologous antiplatelet antibody in the culture system inhibited proplatelet formation, compared with that in controls.     

Culture requirements of Ascaris suum larvae using a stationary multi-well system: Increased survival,development and growth with cholesterol

Ascaris suum third-stage larvae (L₃) were converted from infected rabbits and cultured in a stationary multi-well plate system. Several different culture media including Mediuim 199, NCTC-135, Minimum Essential Medium-Eagle, Dulbecco's Modified Eagle's Medium, RPMI 1640, McCoy's 5A and Neuman-Tytell mdium were tested to determine which was best for overall larval survival, development and growth. Larvae developed from L₃ to fourth-stage (L₄) in all media tested, but larval survival and the yield and growth of L. were superior in RPMI 1640. Pyruvate is an important medium component because its addition to RPMI 1640 enhanced the yield and growth of L₄ while its removal from DMEM reduced larval survival and the yield and growth of L₄. Cholesterol markedly enhanced larval survival and the yield and growth of L₄ when added to RPMI 1640 as either a soluble supplement in Tween 80 or to a lesser degree, as a component of liposomes. The multi-well culture plate system is a convenient method for determining the effects of different media and changes in media composition on larvae in vitro.     

附红细胞体体外杀灭试验

以RPMI 1640为基础培养液,加入20%犊牛血清。选用贝尼尔、红弓、华蜍素、特效米先、附红净、大蒜素、土霉素等药物,在5%CO2的气体条件下对猪附红细胞体和牛附红细胞体进行了体外杀灭试验。结果表明,贝尼尔对猪附红细胞体杀灭作用最好,华蜍素次之;红弓对牛附红细胞体杀灭作用最好,特效米先次之。其余药物杀灭作用不明显。