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171.
172.
牛体外受精胚胎衍生干细胞能力影响因素的研究 总被引:1,自引:0,他引:1
以牛卵巢卵母细胞体外受精获取囊胚期胚胎,比较体外受精牛胚胎不同获取内细胞团的方法和不同培养液对体外培养胚胎干细胞能力的影响。结果表明,囊胚期胚胎不除去透明带而直接培养产生胚胎干细胞,初次克隆率为55%;胰酶法去透明带分离的内细胞团(ICM)培养产生胚胎干细胞,初次克隆率为90%。免疫外科法去透明带分离的ICM在4种不同的培养液中,初次克隆率分别为16.4%、11.5%、21.8%、18.2%,但是其分离的ICM经传代后形成的衍生细胞不易发生分化,经过4~6代的传代,仍然保持其完整的形态,说明免疫外科法是一种理想的牛ICM分离法,培养液为D20+LIF(40ng/mL)可用于牛胚胎干细胞培养。 相似文献
173.
采集家兔自然交配后96h的早期囊胚,以低糖DMEM+150mL/L胎牛血清+0.1mmol/L非必需氨基酸+100IU/mL青霉素+100IU/mL链霉素为基础培养基,比较了不同胚胎处理方法、不同饲养层以及培养液中添加不同成分对兔早期囊胚贴壁和增殖的影响,以完善兔胚胎干细胞的建系方法。结果表明,以胚胎分割法和链霉蛋白酶-E(proteinaseE)处理掉黏蛋白及部分透明带的胚胎容易贴壁和增殖;在小鼠成纤维细胞饲养层和兔胎儿成纤维细胞饲养层上,胚胎的脱带率差别不大,但在小鼠成纤维细胞饲养层上贴壁率明显提高,且贴壁后内细胞团增殖较快;添加胰岛素、白血病抑制因子和伊巯基乙醇均利于抑制ES的分化和促进内细胞团的增殖。 相似文献
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Ki-Soo Park Yong-Soon Lee Kyung-Sun Kang 《Journal of veterinary science (Suw?n-si, Korea)》2006,7(4):343-348
Mesenchymal stem cells (MSCs) have the capabilities for self-renewal and differentiation into cells with the phenotypes of bone, cartilage, neurons and fat cells. These features of MSCs have attracted the attention of investigators for using MSCs for cell-based therapies to treat several human diseases. Because bone marrow-derived cells, which are a main source of MSCs, are not always acceptable due to a significant drop in their cell number and proliferative/differentiation capacity with age, human umbilical cord blood (UCB) cells are good substitutes for BMCs due to the immaturity of newborn cells. Although the isolation of hematopoietic stem cells from UCB has been well established, the isolation and characterization of MSCs from UCB still need to be established and evaluated. In this study, we isolated and characterized MSCs. UCB-derived mononuclear cells, which gave rise to adherent cells, exhibited either an osteoclast or a mesenchymal-like phenotype. The attached cells with mesenchymal phenotypes displayed fibroblast-like morphologies, and they expressed mesenchym-related antigens (SH2 and vimentin) and periodic acid Schiff activity. Also, UCB-derived MSCs were able to transdifferentiate into bone and 2 types of neuronal cells, in vitro. Therefore, it is suggested that the MSCs from UCB might be a good alternative to bone marrow cells for transplantation or cell therapy. 相似文献
178.
ZHOU Dun-hua WEI Jing HUANG Shao-liang FANG Jian-pei BAO Rong WU Yan-feng LI Yang 《园艺学报》2006,22(8):1606-1609
AIM:To study the isolation,expansion and purification of mesenchymal stem cells (MSCs) from human umbilical cord blood (UCB),and investigate some biological identities of MSCs.METHODS:(1) MSCs of UCB,adult bone marrow (BM) and fetus BM were isolated by centrifugation with Ficoll,and the different kinds of MSCs were observed everyday.(2) Surface markers of MSCs were identified by flow cytometry.(3) The level of HGFs (TPO,SCF,FLT-3L,IL-6) secreted by different sources of MSCs was checked by ELISA method.RESULTS:(1) No difference in morphology of the colonies between UCB MSCs and BM MSCs was observed.However,the mononuclear cells needed in culture of UCB MSCs was about 3 times more than that in culture of BM MSCs.The times of UCB MSCs colony formation and confluencing were longer than that of BM in primary culture.(2) After passaged,there was no significant difference in the proliferation rates of 3 kinds of MSCs.Only 4 of 15 UCB samples contained a homogeneous population of MSCs.(3) UCB MSCs shared the same markers with BM MSCs.Neither hematopoietic marker nor immunologic recognition antigens were expressed.(4) The level of hematopoietic growth factors (HGFs) secreted by 3 kinds of MSCs was similar.CONCLUSIONS:(1) MSCs were isolated from UCB,but the amount of MSCs in UCB was smaller than that in BM,and just seldom samples of UCB contained homogeneous MSCs.(2) MSCs from UCB and BM shared the same biological characteristics,such as proliferation ability,surface markers,immunophenotypes and HGFs secretion. 相似文献
179.
地高辛标记cDNA探针检测苹果茎痘病毒 总被引:3,自引:0,他引:3
Partial sequence(314 bp) of ASPV was cloned and used as a probe labelled with digoxigenin-11dUTP. The total RNA extracted from samples with Apple stem pitting virus and a series of dilutions of plasmid with ASPV-cDNA were detected by dot blot hybridization. The results showed that the probe was sensitive and specific. The probe couldn't hybridize with total RNA of Apple stem grooving virus, Apple mosaic virus and Apple chlorotic leaf spot virus samples as well as negative control, only hybridized with that extracted from dormant shoot infected with ASPV. The sensitivity for detection of plasmid contained ASPV-cDNA was 1.64 μg. 相似文献
180.
AIM: To study chromosome aberration due to ethidium bromide (EB),a heterocyclic organic compound and an organic fluorescence dye commonly used in biochemical experiment,and to help further understanding the molecular mechanism of tumor or cancer induced by EB and other heterocyclic organic compounds.METHODS: The toxicity action of EB was evaluated from three aspects including DNA,chromosome and embryo stem cells (ESCs) using atomic force microscopy (AFM),and thereinto,the morphology structural difference of ESCs treated with two EB doses was also valuated.RESULTS: The morphological structures of DNA,chromosome and ESCs were dramatically damaged.The average height of DNA decreased 0.5 nm;chromosomal arms were ruptured from centromere location;molecules of cellular membrane congregated and loop-like structure formed,and ES cell masses were collapsed and became dead after large EB doses treatment and mesh-like morphological structure was discernable.CONCLUSION: The toxicity action of EB is strong and destroys the surface structure of DNA and chromosome.EB induces structural aberration of ES cellular membrane and cell death.The results indicate that the action of EB is externalized at gene level and cell level,which is important to study the carcinogenicity of EB. 相似文献