一、方法根据1999年lutz的方法，做了一些改进，培养DC很多人用的是高浓度的细胞，高浓度的因子，但是这样产量不高，我每一次分离一只小鼠的骨髓，大约可以得到107个细胞，分成10盘，每盘10 ml培养基，用低浓度的细胞因子，这样可以大大提高产量，每次能培养得到undefined107个DC左右，流式细胞术检测：细胞纯度应该大于90%，这个方法还是很好用的。二、具体步骤1. C57Bl/6小鼠2. 处死小鼠，75%酒精浸泡10 min。3. 解剖取出小鼠股 ...
细胞培养（特别是传代细胞）被支原体污染是个世界性问题，在细胞培养中支原体感染发生率达到63%,只要有一个具有增殖能力的支原体就能引起培养体系的污染,支原体则像慢性毒药一般，杀细胞于无形中，据统计，超过25%的细胞系中感染了支原体，研究者又不能通过可视法对其进行检测，而且它对细胞的生长率影响较小，也不易引起注意，所以研究者并不知晓。支原体污染发生后危害巨大，能改变细胞的DNA,RNA及蛋白表达，改变细胞、篡改数据，造成 ...
聚合酶链式反应(Polymerase Chain Reaction，PCR)可对特定核苷酸片断进行指数级的扩增。在扩增反应结束之后，我们可以通过凝胶电泳的方法对扩增产物进行定性的分析，也可以通过放射性核素掺入标记后的光密度扫描来进行定量的分析。无论定性还是定量分析，分析的都是PCR终产物。但是在许多情况下，我们所感兴趣的是未经PCR信号放大之前的起始模板量。例如我们想知道某一转基因动植物转基因的拷贝数或者某一特定基 ...
Out of sight, out of mind.First off, the obligatory warning. Don't try this at home, kids! In the hands of inexperienced laymen, the method I am about to describe will inevitably be a disaster. Like, fatal-type disaster. Consider yourself forewarned.That said, I have to admit that Jillian was truly one to die ...
Fluorescent-based technologies offer opportunities for developing new assays for detection, quantification, and characterization of viral isolates. According to the intrinsic characteristics of fluorescent-based tools (high specificity, sensitivity, and relia ...
miRNAs have emerged as key regulators of gene expression in both plants and animals. These small (generally 21–22 nt) RNA molecules, originated from primary “hairpin” transcripts, can induce translational suppression or direct mRNA cleavage. Similar to regular mRNAs, the expression of ...
Non-invasive microelectrode ion flux measuring (the MIFE system) allows concurrent quantification of net fluxes of several ions with high spatial (several μm) and temporal (ca 5 s) resolution. Over the last 10 years, the MIFE system has been widely used to study various aspects of salt stress sig ...
Nearly all signal transduction pathways lead to regulation of gene expression by controlling specific transcription factors (TFs). Chromatin immunoprecipitation (ChIP) is a powerful method for studying TF–DNA interactions in vivo. To identify all binding sites of a TF in the genome, t ...
Chimeric REpressor gene Silencing Technology (CRES-T) is a useful tool for functional analysis of plant transcription factors. In this system, a chimeric repressor that is produced by fusion of a transcription factor to the plant-specific EAR-motif repression domain (SRDX) suppres ...
Chloroplasts are metabolically important organelles that perform many essential functions within plant cells. The chloroplasts can be subdivided into six distinct sub-compartments to which a protein may be ultimately targeted. These sub-compartments are defined as the outer e ...
Soil salinity reduces the ability of plants to take up water, and this quickly causes reductions in the rate of cell expansion in growing tissues. The slower formation of photosynthetic leaf area in turn reduces the flow of assimilates to the meristematic and growing tissues of the plant. Later, salt ...
Our group recently provided evidence for the presence of neural stem cells and/or progenitor cells in the adult human spinal cord. In this chapter, we review materials and methods to harvest high-quality samples of thoracolumbar, lumbar, and sacral adult human spinal cord from brain-dead pa ...
Adult neural precursor cells are an essential part of the brain, and a focus of two decades of intense research (Ming and Song, Neuron 70:687–702, 2011). Even though adult human stem/progenitor cells have been identified early on (Kirschenbaum et al., Cereb Cortex 4:576–589, 1994; Eriksson et al., Nat M ...
NSCs have been demonstrated to be very useful in grafts into the mammalian central nervous system to investigate the exploitation of NSC for the therapy of neurodegenerative disorders in animal models of neurodegenerative diseases. To push cell therapy in CNS on stage of clinical applica ...
Whereas neural stem cells and their niches have been extensively studied in the brain, little is known on these cells, their environment and their function in the adult spinal cord. Adult spinal cord neural stem cells are located in a complex niche surrounding the central canal and these cells expr ...
It is now well established that a resident population of neural precursor cells continues to generate new neurons in the adult hippocampus throughout life. Numerous studies have suggested that these newborn neurons preferentially participate in the functional hippocampal circ ...
Due to the complexity of the NSC niche organization, the lack of specific NSC markers and the difficulty of long-term tracking these cells and their progeny in vivo the functional properties of the endogenous NSCs remain largely unexplored. These limitations have led to the development of met ...
In vitro studies of neural progenitors isolated from the developing mouse have provided important insights into intrinsic and extrinsic pathways that control their behavior. However, use of primary cultures or neurospheres established from fetal tissues in cell population-ba ...
The propagation of neural precursors in culture is an essential tool for the study of the signaling matrix that regulates their proliferation, self-renewal, and generation of terminally differentiated progeny. Neural precursors can be expanded in vitro using both adherent and non-a ...
The olfactory mucosa, located in the nasal cavity, is the only nervous tissue that is exposed to the external environment and easily accessible in every living individual. In addition, this organ is home of a continuing neurogenesis that is sustained by a large population of stem cells. Here, we desc ...
