动物乳腺干细胞的研究进展

乳腺干细胞具有产生所有类型乳腺细胞的能力。在乳腺组织生长发育过程中,乳腺干细胞对于动物青春期、妊娠期、泌乳期和泌乳衰退期的乳腺生长与重建等具有重要意义。体外培养的乳腺干细胞是研究乳腺细胞增殖、分化、生存和凋亡等信号通路的理想模型,同时在转基因乳腺生物反应器方面也具有重要的应用前景。因此,乳腺干细胞的研究对于乳腺发育的认知,乳腺癌的治疗及农业生产具有重要意义。作者针对乳腺干细胞的筛选、定位和体外培养方法的研究现状进行了系统的综述,并对未来的研究方向与应用前景加以展望。     

奶牛干细胞多能性基因Nanog和Lin28克隆分析及其逆转录病毒表达载体的构建与包装

以50~60日龄的Holstein奶牛胎儿原始生殖嵴组织为材料,通过RT-PCR的方法克隆奶牛Nanog与Lin28基因开放性阅读框(ORF)全序列。序列全长分别为903 bp和618 bp。所克隆的奶牛Nanog与Lin28基因序列与GenBank中已经发表的参考序列进行比对同源性均为100%。将扩增的基因片段经过酶切后亚克隆到反转录病毒载体pMSCVneo的多克隆位点,经过酶切、PCR和测序鉴定正确后,应用lipofectamine2000介导转染PT67包装细胞,经过14 d G418连续抗性筛选,挑选抗性集落扩大培养。收集病毒上清经过RT-PCR与透射电镜检测,并用NIH3T3细胞测定病毒滴度。结果表明,病毒上清中含有逆转录病毒颗粒,Nanog与Lin28基因在筛选后的包装细胞PT67细胞中均有转录,病毒上清感染滴度值分别达到7.5×107cfu/mL和9.19×107cfu/mL。结果表明,本试验已经成功克隆出奶牛Nanog与Lin28基因开放性阅读框全序列,并建立高效、稳定产生包含有pMSCV-Nanog与pMSCV-Lin28质粒的感染性病毒颗粒的包装细胞株,为进一步产生奶牛诱导性多能干细胞(Induced pluripotentstem cells,iPSCs)奠定基础。     

Mammary stem cells： expansion and animal productivity

Identification and characterization of mammary stem cells and progenitor cells from dairy animals is important in the understanding of mammogenesis, tissue turnover, lactation persistency and regenerative therapy. It has been realized by many investigators that altered lactation, long dry periods （non-milking period between two consecutive lactation cycles）, abrupt cessation of lactation （common in water buffaloes） and disease conditions like mastitis, greatly reduce milk yield thus render huge financial losses within the dairy sector. Cellular manipulation of specialized cell types within the mammary gland, called mammary stem cells （MaSCs）/progenitor cells, might provide potential solutions to these problems and may improve milk production. In addition, MaSCs/progenitor cells could be used in regenerative therapy against tissue damage caused by mastitis. This review discusses methods of MaSC/progenitor cell manipulation and their mechanisms in bovine and caprine animals. Author believes that intervention of MaSCs/progenitor cells could lessen the huge financial losses to the dairy industry globally.     

Identification of Suspected Stem/progenitor Cells in Bovine Mammary Epithelial Cells Culture System

To develop the potential function of dairy cow mammary stem cells (DCMECs) in regulation of lactation,we identify putative DCMECs which were BrdU label retaining epithelial cells,at the same time,analysis the location of two new mammary stem cells molecular marks FNDC3B and PROCR to verify the feasibility of them to indicate DCMECs.The mRNA levels of prolactin,growth hormone,insulin-like growth factor-1 and their receptors were detected along with cell passage by Real-time quantitative PCR.The results showed that the proportion of BrdU label-retaining epithelial cells was nearly 0.4% after 25 d continuous culture (passaged 4 times) and few cells were positive for FNDC3B or PROCR.Moreover,we observed the BrdU labelled epithelial cells by asymmetric division.The mRNA levels of prolactin,growth hormone,insulin-like growth factor-Ⅰ and their receptors in primary and passage cells were extremely significant difference(P<0.01).DCMECs would rapidly lose some physiological characteristics and the ability of milk synthesis when not under the condition of induction of lactation differentiation,but a certain percentage of mammary stem/progenitor cells will be retained,whose potential effects on the regulation of lactation and mammary acinar remodeling were worthy of attention.     

一株从片烟中分离的纤维素酶产生菌B.Pumilus培养条件优化研究

[目的]优化短小芽孢杆菌(B.Pumilus)菌株TX3产纤维素酶条件,为其降解片烟中所含纤维素提供理论依据.[方法]应用单因子试验考察碳源种类、烟梗粉末添加量、硫酸铵浓度和pH对菌株TX3发酵产纤维素酶的影响,在此基础上选取烟梗粉末添加量、硫酸铵浓度、pH作为显著因子,采用响应面分析法对此3种显著因子进行优化和分析.[结果]菌株TX3最适产酶条件为发酵初始pH6.93,烟梗粉末添加量1.24％,硫酸铵浓度0.35％,以此条件进行摇瓶发酵72 h,其纤维素酶活力达到9.91 U/ml,比优化前高了31.4％.[结论]烟梗粉末添加量、硫酸铵浓度、pH对菌株TX3产纤维素酶单个及交互影响均显著.     

修枝截干对二球悬铃木叶片光合特性的影响

采用Li-6400便携式光合作用测定系统对修枝截干和自然生长的二球悬铃木中部当年向南枝顶的完全展开叶进行单叶光合特性测定.结果表明,修枝截干和自然生长的二球悬铃木叶片的净光合速率(Pn)日变化均呈双峰曲线,有明显的"午休"现象,最高峰发生在11:00,次高峰在15:00,光强最大时(13:00) Pn最低,即两种生长状态下都受到光抑制.修枝截干二球悬铃木Pn[μmol(CO2)·m-2·s-1]的最高峰为12.16,次高峰为9.21,最低谷为6.47,分别比自然生长高27.06%、69.93%和159.84%,这暗示修枝截干可促进光合作用,减轻光抑制.修枝截干的Pn-光量子通量密度(PPFD)响应可拟合为Pn=12.30(1-1.135 3e-0.030 2PPFD/12.30)(R2 =0.989 1**),自然生长的为Pn=10.92(1-1.130 7e-0.030 5PPFD/10.92)(R2 =0.984 9**),修枝截干的最大光合速率(Pmax)、光补偿点、光饱和点及光合幅度的估算值分别为12.30 μmol(CO2)·m-2·s-1、51.67 μmol·m-2·s-1、1 927.32 μmol·m-2·s-1和1 875.65 μmol·m-2·s-1,分别比自然生长提高12.64%、17.33%、13.85%和13.76%,这意味着修枝截干可提高单叶对光能的利用.修枝截干的Pn-CO2浓度(Ci)响应可拟合为Pn=11.96(1-1.471 6e-0.028 7Ci/11.96)(R2=0.982 4**),自然生长的为Pn=10.70(1-1.465 7e-0.029 2Ci/10.70)(R2 = 0.981 0**),修枝截干的Pmax、CO2补偿点、CO2饱和点及CO2幅度估算值分别为11.96 μmol(CO2)·m-2·s-1、67.54 μmol·mol-1、872.02 μmol·mol-1和804.48 μmol·mol-1,分别比自然生长提高11.78%、-27.44%、0.39%和3.73%,说明修枝截干可增强叶片对低浓度CO2的同化能力.     

半乳糖凝集素1蛋白及其生物学功能

半乳糖凝集素1(Galectin-1)是一种分子质量约为14 ku的β-糖结合蛋白,是动物凝集素家族的成员之一。作为多种癌症的诊断指标,治疗癌症的新突破口,对Galectin-1的研究备受关注。此外,Galectin-1分布广泛,与多种正常生物功能相关,如细胞生长、神经修复、血管再生、软骨形成等。现阶段,关于Galectin-1在鹿茸再生中的研究很少,但鹿茸再生中许多过程与Galectin-1的功能高度相关。与肿瘤同样高速生长且高表达Galectin-1的鹿茸组织并不发生癌变,这可能对癌症的研究有所启发。为了寻找治疗癌症的新方法,解释鹿茸再生机制,了解Galectin-1蛋白在肿瘤和鹿茸中的功能研究进展至关重要,文章就其进行了综述。