分化抑制物对动物胚胎干细胞克隆率的影响

胚胎干细胞是从动物胚胎内细胞团或原始生殖细胞分离的具有全能性的细胞，在遗传学、发育生物学和细胞工程领域具有广阔的应用前景。抑制胚胎干细胞分化和促进胚胎干细胞增殖是克隆胚胎干细胞的关键，本文介绍了饲养层、条件培养基、分化抑制因子对哺乳动物胚胎干细胞分离与克隆的影响，并探讨了白血病抑制因子影响胚胎干细胞克隆的机理，认为在饲养层中添加分化抑制因子能有效提高胚胎干细胞克隆率。     

细胞外环境中分化抑制物对动物胚胎干细胞克隆率的影响

介绍了饲养层、条件培养基、分化抑制因子对哺乳动物胚胎干细胞分离与克隆的影响，并探讨了白血病抑制因子影响胚胎干细胞克隆的机理。     

昆明小白鼠胚胎干细胞分离培养的影响因素

为了研究影响胚胎干细胞(ESC)体外分离克隆的因素,试验采用3~5日龄大鼠心肌细胞制成心肌细胞条件培养基(RH-CM),比较了4种培养体系对小鼠胚胎干细胞体外培养的影响,并分析比较了不同饲养层对胚胎干细胞分离克隆造成的影响。结果表明:添加重组鼠白血病抑制因子和心肌细胞条件培养基在胚胎贴壁率、内细胞团(ICM)形成率和原代胚胎干细胞集落形成率以及抑制分化的过程中起到的作用显著优于未添加任何诱导成分的培养基;作为饲养层,小鼠胚胎成纤维细胞(MEF)分离培养胚胎干细胞的效果优于小鼠睾丸支持细胞(MS)。     

小鼠胚胎干细胞培养相关影响因素的研究

本实验主要探讨饲养层、白血病抑制因子(LIF)、水、血清对小鼠胚胎干细胞培养的影响。结果表明:饲养层细胞在小鼠胚胎干细胞的分化抑制过程中有一定作用,而起主导作用的是外源白血病抑制因子;;经亚沸处理的MilliQ水,能显著提高小鼠胚胎干细胞保持未分化状态的能力;;血清的品质对小鼠胚胎干细胞的培养效果影响显著(p<0.05)。     

类ES细胞持续培养的上清液对小鼠囊胚ICM团形成的影响

胚胎干细胞是一种存在于动物早期胚胎中、具有分化全能性的细胞。维持这种细胞的分化全能性与不断扩增的前提条件是：培养环境中必须存在抑制其分化、促进其生长的因子。因此，寻找胚胎干细胞促生长与抑分化因子，是胚胎干细胞研究的重要内容之一。1988年Williams等人发现了能诱导髓样白血病细胞向正常分化的LIF，也能抑制胚胎干     

从原始生殖细胞中分离克隆胚胎干细胞的研究

作者探讨了从原始生殖细胞（PGCs）中分离克隆胚胎干细胞的优势、用于分离克隆胚胎干细胞的PGCs获取方法及其向胚胎干细胞转化的能力。     

动物胚胎干细胞的研究概况

胚胎干细胞（ES细胞）是从早期胚胎内细胞团（ICM）或原始生殖细胞（PGCs）经体外分化抑制培养分离克隆的,ES细胞在动物克隆、转基因动物生产、细胞工程、组织工程、临床克隆治疗和发育生物学等的研究应用中起着重要的作用.为此,介绍胚胎干细胞的生物学特性,国内外研究进展和研究动态,阐明建立ES细胞系的技术要点以及ES细胞的应用及发展前景.     

影响胚胎干细胞分离克隆的因素分析

本文就胚胎质量和胚胎类型、分化抑制物、培养基、传代时间和消化液等方面对胚胎干细胞分离克隆的影响进行了讨论。     

动物胚胎干细胞的概念及特性

胚胎干细胞是从早期胚胎内细胞团或原始生殖细胞分离和克隆的具有全能性的细胞。其在动物克隆转基因动物的生产、基因结构与功能的研究以及细胞分化空的研究方面具有广泛的应用,对胚胎干细胞的概念特性以及胚胎干细胞特性之间的关系进行了论述。     

胚胎干细胞的研究进展及其应用

胚胎干细胞是当今生物医学领域研究的重点,本文综述了国内外对胚胎干细胞的最新研究成果,以及胚胎干细胞在治疗性克隆、药物研发、发育生物学、转基因动物等方面的应用。     

鸡胚胎干细胞及其应用研究进展

鸡胚胎干细胞是一种多能性干细胞,从X期胚盘分离胚盘细胞或早期鸡胚的生殖嵴分离原始生殖细胞,经体外长期抑制分化培养可得到鸡胚胎干细胞。为维持细胞在培养过程中的未分化状态,需要采用饲养层细胞培养,同时设计合理的培养液配方并添加多种抑制分化或促进增殖的细胞因子。通过碱性磷酸酶活性检测、胚胎表面特异性抗原检测、分化试验及嵌合体试验等方法,可对鸡胚胎干细胞进行准确鉴定。文章主要就鸡胚胎干细胞的分离、培养与鉴定方法的研究进展及其应用前景进行简要综述,为进一步发展更高效的鸡胚胎干细胞培养体系并应用于生产实践提供一定的借鉴。     

小鼠胚胎干细胞向骨骼肌细胞分化的研究进展

近年来,胚胎干细胞的应用越来越广泛,在体外将小鼠胚胎干细胞诱导分化为肌肉细胞,并且利用这些分化得来的肌肉细胞治疗肌肉退行性疾病,一直是胚胎干细胞研究领域的热点,而胚胎干细胞的分化机制更是其中的难点。目前,用于诱导小鼠胚胎干细胞分化为骨骼肌细胞的方法很多,但分化的效率并不是很高,所以研究胚胎干细胞向骨骼肌细胞方向分化的机制显得尤为重要。文章仅就最近几年对小鼠胚胎干细胞向骨骼肌细胞方向分化的一些方法及其机制作一综述。     

Analysis of marker expression in porcine cell lines derived from blastocysts produced in vitro and in vivo

The present study was designed to extensively characterize cell lines derived from porcine blastocysts by several methodical approaches, including morphological observation, cytogenetic analysis, estimation of alkaline phosphatase activity and detection of specific marker expression at the mRNA/protein level. A comparison was made between the properties of cell lines isolated from in vivo- and in vitro-obtained blastocysts. Our results showed that 57.1% of the in vivo-obtained blastocysts attached to the feeder layer and that 33.3% of them started to grow in a monolayer. The percentage of attached in vitro-produced blastocysts was lower (24.6%), and only 6.9% of them started to grow. Outgrowths from the in vitro-produced blastocysts formed mainly trophectoderm or epithelial-like monolayer, whereas the in vivo-obtained blastocysts formed heterogeneous outgrowths that also contained cells with embryonic stem (ES)-like morphology. Detailed analyses showed that the primary outgrowths with ES-like morphology expressed the pluripotency markers OCT-4 and NANOG and revealed intensive alkaline phosphatase staining, while they did not express markers of differentiation. The majority of passaged cells, including those with ES-like morphology, lacked OCT-4 protein and revealed expression of specific differentiation markers (cytokeratin 18, lamins A/C, transferrin, α-fetoprotein and GATA-4), although they still expressed NANOG and exhibited weak alkaline phosphatase activity. Moreover, these cells spontaneously differentiated into neural, fibroblast or epithelial-like cells, even in the presence of leukaemia inhibitory factor. Our results show that complex analysis of markers of pluripotency as well as differentiation markers is necessary for proper interpretation of data in porcine embryonic stem cell studies.     

鸡胚胎干细胞的分离和培养

实验从鸡第X期胚胎中分离胚盘,以鸡成纤维细胞为饲养层,用添加了10%的胎牛血清、2%的鸡血清、2mmol/LL-谷氨酰胺、1mmol/L丙酮酸钠、5.5×10-5mol/Lβ-巯基乙醇、10μL/mL非必需氨基酸以及含1000IU/mL白血病抑制因子(LIF)、10ng/mL碱性成纤维生长因子(bFGF)和5ng/mL干细胞生长因子(SCF)的高糖DMEM对细胞进行培养和传代,可以获得传至5～6代的鸡胚胎干细胞(ES)。通过对传代培养后的鸡ES细胞进行AKP染色鉴定和SSEA-1的鉴定,证实细胞未发生分化,具有胚胎干细胞的特征。同时通过不同分离胚盘方法和不同消化时间的比较得出药勺法提取胚盘简单易行,原代消化5～8min的ES细胞适合于传代培养。     

鱼类胚胎干细胞的研究进展

胚胎干细胞是存在于早期胚胎或原始生殖嵴,具有发育全能性和无限增殖能力的一类细胞。作为实验材料,该类细胞已广泛应用于基因的表达与调控、细胞分化的机制、动物克隆及转基因动物生产等研究领域。与其他哺乳动物胚胎干细胞的研究相比,虽然鱼类胚胎干细胞的研究起步较晚,但近年来该领域的研究进展迅速。本文综述了鱼类胚胎干细胞的分离培养、生物学特性、体外分化能力及嵌合体形成等方面的研究进展。     

Isolation and culture of rabbit primordial germ cells

Primordial germ cells (PGCs) are embryonic precursors of the gametes of adult animals and are considered stem cells of the germline. Since their proliferation in vitro correlates well with the schedule of developmental changes in vivo, they might be interesting research tools for genomic imprinting, germ-cell tumors and fertility. Furthermore, once primordial germ cells are separated and placed on a feeder layer with cytokines, they become cultured pluripotent cell lines called embryonic germ (EG) cells. EG cells share several important characteristics with embryonic stem (ES) cells as they can also contribute to the germ line of chimeras. To investigate the characteristics of PGCs and establish rabbit EG (rEG) cells, we cultured rabbit PGCs (rPGCs) in vitro with various combinations of leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and forskolin on inactivated mouse embryonic fibroblast (MEF) feeder layers. The present study found PGC proliferation in early cultures and induction of rEG-like colonies. These cells expressed pluripotent markers, such as alkaline phosphatase activity, OCT-4, Sox-2 and SSEA-1, in the undifferentiated state; however, the cells did not develop into a teratoma when injected into the kidney capsules of SCID mice, although the restricted differentiation potentials to neural cells were determined via embryoid body formation. From these characteristics and further characterization of the germ stem cell markers Vasa, SCP-1 and SCP-3, we suggested that these were hybrid cells with characteristics somewhere between PGC and EG cells.     

鱼类胚胎干细胞的研究进展

胚胎干细胞是存在于早期胚胎或原始生殖嵴,具有发育全能性和无限增殖能力的一类细胞.作为实验材料,该类细胞已广泛应用于基因的表达与调控、细胞分化的机制、动物克隆及转基因动物生产等研究领域.与其他哺乳动物胚胎干细胞的研究相比,虽然鱼类胚胎干细胞的研究起步较晚,但近年来该领域的研究进展迅速.本文综述了鱼类胚胎干细胞的分离培养、生物学特性、体外分化能力及嵌合体形成等方面的研究进展.