非人灵长类局部脑缺血动物模型研究现状

非人灵长类动物在种系发生上较啮齿类更接近于人类,用来制备局部脑缺血模型可以更好的拟合临床症状和机理。通过对国内外非人灵长类动物局部脑缺血模型的制备方法和应用现状进行收集、分类和述评,展望非人灵长类动物模型的应用前景,尤其是利用低等灵长类动物树鼩研究缺血性中风的优势。     

恒河猴在生殖生物学中的应用进展

目的非人灵长类动物在生殖生物学研究领域是一种非常重要的实验动物。人类利用非人灵长类动物与人的生物学等方面相似的特征,开展了生殖生物学、生理学、药理学、毒理学以及生育调节等方面的研究工作,为生殖生物学基础研究以及人类健康和疾病问题的基础研究和临床前研究提供了理想的动物模型。随着生命科学的发展,对非人灵长类实验动物质量提出了更高的要求,人们认识到实验时,应用健康的优质非人灵长类动物的重要性。本文简要概括了非人灵长类动物恒河猴的生物学特性,阐述了非人灵长类动物恒河猴在生殖生物学中的应用研究。     

通过CRISP刚Cas9和TALENs介导的基因打靶技术获得基因修饰的猴模型

1研究背景 灵长类动物,如猕猴和食蟹猴,在遗传和生理特性上与人类具有很高的相似性,是研究人类疾病、发育和临床前治疗等方面最为理想、有时甚至是唯一的实验动物。通过遗传修饰的方法获得灵长类动物模型对探讨疾病的致病机理和治疗有重大的意义,因此科学家一直在努力构建理想的灵长类动物模型。来自美国、     

非人灵长类动物种质冷冻保存研究进展和展望

非人灵长类动物是生物多样性的重要组成部分,也是生物医学研究的珍贵实验动物,然而,由于人类活动、栖息地破坏、狩猎和遗传隔离等原因,许多非人灵长类动物的野生种群数量急剧下降,甚至处于灭绝的边缘。种质冷冻保存对拯救非人灵长类动物和保存遗传物质资源具有重要意义。本文综述了新大陆猴、旧大陆猴和巨猿等类群动物精子、卵子、胚胎和性腺组织等种质冷冻保存的研究进展,介绍了狨猴、松鼠猴、恒河猴、食蟹猴和黑猩猩等种质冷冻保存的主要方法,并对未来种质冷冻保存的研究方向进行了讨论。     

性早熟动物造模研究进展

性早熟是儿童常见内分泌疾病，动物实验在研究性早熟的发病机制和药物治疗中必不可少，因此建立合适的动物模型是研究性早熟的必要前提。啮齿类(如大鼠)与非人灵长类动物的青春期发育过程与人类近似，性腺发育指征明确，是性早熟模型常用动物。根据研究内容不同出现了达那唑、N-甲基-DL-天冬氨酸、雌二醇、高脂饲料、不良生活条件、光照及褪黑素、锰暴露、环境内分泌干扰物、颅脑辐射、脑内药物注射等多种性早熟造模方法。目前多采用阴道开口时间、阴道涂片及动情周期观察、血清性激素水平测定、下丘脑基因检测、性器官检测等多种指标对性早熟动物实验进行评估。     

非人灵长类动物基因编辑技术的应用及挑战

建立有效的动物模型是研究人类疾病演进、开发新型治疗手段的重要方法。非人灵长类动物在进化发育、生理生化及病理方面和人类最接近,是研究人类疾病的理想动物模型。随着基因编辑技术的发展,研究者已经成功建立了多种模仿人类疾病的非人灵长类动物模型。但是CRISPR/Cas9的脱靶效应、嵌合突变以及基因敲入效率较低等突出问题也逐渐引起重视。本文综述了基因编辑技术在建立非人灵长类动物模型中的应用现状,提出了目前亟需解决的难点和应对策略,以期为高效、准确构建非人灵长类动物模型提供借鉴与参考。     

非人灵长类动物在医学科学实验中的应用

非人灵长类动物在亲缘关系上和人最接近,与人类的遗传物质有75%～98.5%的同源性,在组织结构、免疫、生理和代谢等方面与人类高度近似,是极其珍贵的实验动物,其应用价值远超过其他种属的实验动物。本文就非人灵长类和人类之间的进化关系和目前的使用情况,及其在毒理学、传染病、神经科学、生殖生物学、胎儿发育和衰老等医学科学实验中的应用等方面的内容做了简要的介绍。     

非人灵长类胚胎工程研究进展

由于非人灵长类动物与人类在进化上的亲缘关系和生理特征上的高度相似,它们在人类传染病、生殖、发育、代谢、神经和衰老等研究领域具有重要的应用价值。现对非人灵长类动物的常见胚胎工程技术进行综述,包括非人灵长类动物的配子收集、体外受精和胚胎移植等;同时,还结合基因组编辑技术的发展,简要介绍了近年来胚胎工程技术在非人灵长类动物基因编辑方面的应用。     

通过CRISPR/Cas9和TALENs介导的基因打靶技术获得基因修饰的猴模型

<正>1研究背景灵长类动物,如猕猴和食蟹猴,在遗传和生理特性上与人类具有很高的相似性,是研究人类疾病、发育和临床前治疗等方面最为理想、有时甚至是唯一的实验动物[1]。通过遗传修饰的方法获得灵长类动物模型对探讨疾病的致病机理和治疗有重大的意义,因此科学家一直在努力构建理想的灵长类动物模型。来自美国、日本和中国的科学家们先后获得了利用逆转录病毒或慢病毒介导的转基因猴[2-5],但这些通过外源基因的随机插入和过量表达来实现对基因组的修饰,无法对确定的目的基因进行修饰和控制,所获得的动物模型具有一定的局限性和不确定性。最近发展起来的锌指核酸酶(zinc-finger nucleases,ZFNs)、转录激活因子样     

非人灵长类动物精子冻存技术的研究进展

随着生物医学技术的发展,应用非人灵长类动物模型进行基础科学研究日益广泛。与此同时,由于栖息地破坏、狩猎和基因隔离,许多非人灵长类动物濒临灭绝。因此,改进非人灵长类动物精子冻存技术对物种遗传资源的保藏具有重要的意义。本文概述了非人灵长类动物的精液特征,介绍了精液液化和冷冻精子质量评估的方法,分析了冷冻保护剂、冷冻稀释液及冷冻方法等因素对精子冻存效果的影响,总结了目前非人灵长类精子冷冻常用的冷冻保存液和冷冻方法,并对相关精子参数进行了比较,同时探讨了非人灵长类动物精子冻存研究面临的困境,并提出了可行的方案。总之,本文综述了近年来非人灵长类动物精子冻存的重要研究成果,对开发新的冷冻保护剂及改进冷冻技术具有一定的参考价值。     

Primate stem cells: bridge the translation from basic research to clinic application

A growing body of literature has shown that stem cells are very effective for the treatment of degenerative diseases in rodents but these exciting results have not translated to clinical practice. The difference results from the divergence in genetic, metabolic, and physiological phenotypes between rodents and humans. The high degree of similarity between non-human primates(NHPs) and humans provides the most accurate models for preclinical studies of stem cell therapy. Using a NHP model to understand the following key issues, which cannot be addressed in humans or rodents, will be helpful for extending stem cell applications in the basic science and the clinic. These issues include pluripotency of primate stem cells, the safety and efficiency of stem cell therapy, and transplantation procedures of stem cells suitable for clinical translation. Here we review studies of the above issues in NHPs and current challenges of stem cell applications in both basic science and clinical therapies. We propose that the use of NHP models, in particular combining the serial production and transplantation procedures of stem cells is the most useful for preclinical studies designed to overcome these challenges.     

Transgenic nonhuman primate models for human diseases: approaches and contributing factors

Nonhuman primates (NHPs) provide powerful experimental models to study human development,cognitive functions and disturbances as well as complex behavior,because of their genetic and physiological simi...     

Long-term Continuous EEG Monitoring in Small Rodent Models of Human Disease Using the Epoch Wireless Transmitter System

Many progressive neurologic diseases in humans, such as epilepsy, require pre-clinical animal models that slowly develop the disease in order to test interventions at various stages of the disease process. These animal models are particularly difficult to implement in immature rodents, a classic model organism for laboratory study of these disorders. Recording continuous EEG in young animal models of seizures and other neurological disorders presents a technical challenge due to the small physical size of young rodents and their dependence on the dam prior to weaning. Therefore, there is not only a clear need for improving pre-clinical research that will better identify those therapies suitable for translation to the clinic but also a need for new devices capable of recording continuous EEG in immature rodents. Here, we describe the technology behind and demonstrate the use of a novel miniature telemetry system, specifically engineered for use in immature rats or mice, which is also effective for use in adult animals.     

慢病毒载体感染成年食蟹猴骨髓间充质干细胞

骨髓间充质干细胞(Mesenchymal stem cells,MSCs)具有增殖和多向分化潜能,临床应用广泛,近年来备受关注。另一方面,MSCs易于转导和表达外源基因,是理想的基因工程细胞。非人灵长类(NHPs)和人类具有非常相近的遗传背景,NHPs模型在评价药物疗效和移植治疗等方面具有不可替代的价值。本研究采用密度梯度离心法分离成年食蟹猴骨髓单核细胞(Marrow mononuclear cells,MNCs),贴壁培养MSCs。同时构建表达绿色荧光蛋白(Green fluorescent protein,GFP)的慢病毒载体,感染成年食蟹猴MSCs。结果显示,体外培养的成年食蟹猴MSCs均感染猴泡沫病毒(Simian foamy virus,SFV),体外培养成年食蟹猴MSCs必须添加抗病毒药物Tenofovir。但由于食蟹猴MSCs感染SFV,以及培养中添加了抗病毒药物Tenofovir,慢病毒载体的感染效率明显降低(10%)。本研究通过停用抗病毒药,在细胞复苏后6d转染慢病毒,可大幅提高慢病毒的感染效率(50%)。为成年食蟹猴MSCs作为基因工程细胞应用于实验和临床研究提供了技术保证。     

Pioglitazone Increases Whole Body Insulin Sensitivity in Obese,Insulin-Resistant Rhesus Monkeys

Hyperinsulinemic-euglycemic clamps are considered the "gold standard" for assessing whole body insulin sensitivity. When used in combination with tracer dilution techniques and physiological insulin concentrations, insulin sensitization can be dissected and attributed to hepatic and peripheral (primarily muscle) effects. Non-human primates (NHPs), such as rhesus monkeys, are the closest pre-clinical species to humans, and thus serve as an ideal model for testing of compound efficacy to support translation to human efficacy. We determined insulin infusion rates that resulted in high physiological insulin concentrations that elicited maximal pharmacodynamic responses during hyperinsulinemic-euglycemic clamps. These rates were then used with [U-¹³C]-D-glucose, to assess and document the degrees of hepatic and peripheral insulin resistance between healthy and insulin-resistant, dysmetabolic NHPs. Next, dysmetabolic NHPs were treated for 28 days with pioglitazone (3 mg/kg) and again had their insulin sensitivity assessed, illustrating a significant improvement in hepatic and peripheral insulin sensitivity. This coincided with a significant increase in insulin clearance, and normalization of circulating adiponectin. In conclusion, we have determined a physiological clamp paradigm (similar to humans) for assessing glucose turnover in NHPs. We have also demonstrated that insulin-resistant, dysmetabolic NHPs respond to the established insulin sensitizer, pioglitazone, thus confirming their use as an ideal pre-clinical translational model to assess insulin sensitizing compounds.     

Old world nonhuman primate models of type 2 diabetes mellitus

Type 2 diabetes mellitus is a major health problem of increasing incidence. To better study the pathogenesis and potential therapeutic agents for this disease, appropriate animal models are needed. Old World nonhuman primates (NHPs) are a useful animal model of type 2 diabetes; like humans, the disease is most common in older, obese animals. Before developing overt diabetes, NHPs have a period of obesity-associated insulin resistance that is initially met with compensatory insulin secretion. When either a relative or absolute deficiency in pancreatic insulin production occurs, fasting glucose concentrations begin to increase and diabetic signs become apparent. Pathological changes in pancreatic islets are also similar to those seen in human diabetics. Initially there is hyperplasia of the islets with abundant insulin production typically followed by replacement of islets with islet-associated amyloid. Diabetic NHPs have detrimental changes in plasma lipid and lipoprotein concentrations, lipoprotein composition, and glycation, which may contribute to progression of atherosclerosis. As both the prediabetic condition (similar to metabolic syndrome in humans) and overt diabetes become better defined in monkeys, their use in pharmacological studies is increasing. Likely due to their genetic similarity to humans and the similar characteristics of the disease in NHPs, NHPs have been used to study recently developed agonists of the peroxisome proliferators-activated receptors. Importantly, agonists of the different receptor subclasses elicit similar responses in both humans and NHPs. Thus, Old World NHPs are a valuable animal model of type 2 diabetes to study disease progression, associated risk factors, and potential new treatments.     

Safety testing of monoclonal antibodies in non-human primates: Case studies highlighting their impact on human risk assessment

Monoclonal antibodies (mAbs) are improving the quality of life for patients suffering from serious diseases due to their high specificity for their target and low potential for off-target toxicity. The toxicity of mAbs is primarily driven by their pharmacological activity, and therefore safety testing of these drugs prior to clinical testing is performed in species in which the mAb binds and engages the target to a similar extent to that anticipated in humans. For highly human-specific mAbs, this testing often requires the use of non-human primates (NHPs) as relevant species. It has been argued that the value of these NHP studies is limited because most of the adverse events can be predicted from the knowledge of the target, data from transgenic rodents or target-deficient humans, and other sources. However, many of the mAbs currently in development target novel pathways and may comprise novel scaffolds with multi-functional domains; hence, the pharmacological effects and potential safety risks are less predictable. Here, we present a total of 18 case studies, including some of these novel mAbs, with the aim of interrogating the value of NHP safety studies in human risk assessment. These studies have identified mAb candidate molecules and pharmacological pathways with severe safety risks, leading to candidate or target program termination, as well as highlighting that some pathways with theoretical safety concerns are amenable to safe modulation by mAbs. NHP studies have also informed the rational design of safer drug candidates suitable for human testing and informed human clinical trial design (route, dose and regimen, patient inclusion and exclusion criteria and safety monitoring), further protecting the safety of clinical trial participants.     

Lineage selection and the evolution of multistage carcinogenesis

A wide array of proto-oncogenes and tumour suppressor genes are involved in the prevention of cancer. Each form of cancer requires mutations in a characteristic group of genes, but no single group controls all cancers. This lack of generality shows that the control of cancer is not an ancient, fixed property of cells. By contrast, it supports a dynamic evolutionary model, whereby genetic controls over unregulated cell growth are recruited independently through evolutionary time in different tissues within different taxa. The complexity of this genetic control can be predicted from a population genetic model of lineage selection driven by the detrimental fitness effects of cancer. Cancer occurs because the genetic control of cell growth is vulnerable to somatic mutations (or 'hits'), particularly in large, continuously dividing tissues. Thus, compared to small rodents, humans must have evolved more complex genetic controls over cell growth in at least some of their tissues because of their greater size and longevity; an expectation relevant to the application of mouse data to humans. Similarly, the 'two-hit' model so successfully applied to retinoblastoma, which originates in a small embryonic tissue, is unlikely to be generally applicable to other human cancers; instead, more complex scenarios are expected to dominate, with complexity depending upon a tissue's size and its pattern of proliferation.     

Characterization of Regional Left Ventricular Function in Nonhuman Primates Using Magnetic Resonance Imaging Biomarkers: A Test-Retest Repeatability and Inter-Subject Variability Study

Pre-clinical animal models are important to study the fundamental biological and functional mechanisms involved in the longitudinal evolution of heart failure (HF). Particularly, large animal models, like nonhuman primates (NHPs), that possess greater physiological, biochemical, and phylogenetic similarity to humans are gaining interest. To assess the translatability of these models into human diseases, imaging biomarkers play a significant role in non-invasive phenotyping, prediction of downstream remodeling, and evaluation of novel experimental therapeutics. This paper sheds insight into NHP cardiac function through the quantification of magnetic resonance (MR) imaging biomarkers that comprehensively characterize the spatiotemporal dynamics of left ventricular (LV) systolic pumping and LV diastolic relaxation. MR tagging and phase contrast (PC) imaging were used to quantify NHP cardiac strain and flow. Temporal inter-relationships between rotational mechanics, myocardial strain and LV chamber flow are presented, and functional biomarkers are evaluated through test-retest repeatability and inter subject variability analyses. The temporal trends observed in strain and flow was similar to published data in humans. Our results indicate a dominant dimension based pumping during early systole, followed by a torsion dominant pumping action during late systole. Early diastole is characterized by close to 65% of untwist, the remainder of which likely contributes to efficient filling during atrial kick. Our data reveal that moderate to good intra-subject repeatability was observed for peak strain, strain-rates, E/circumferential strain-rate (CSR) ratio, E/longitudinal strain-rate (LSR) ratio, and deceleration time. The inter-subject variability was high for strain dyssynchrony, diastolic strain-rates, peak torsion and peak untwist rate. We have successfully characterized cardiac function in NHPs using MR imaging. Peak strain, average systolic strain-rate, diastolic E/CSR and E/LSR ratios, and deceleration time were identified as robust biomarkers that could potentially be applied to future pre-clinical drug studies.     

睡眠研究中相关动物选择的进展

在睡眠研究使用的各种实验动物中,除了常用的啮齿类动物,还有猫、果蝇、猴和斑马鱼等.啮齿类动物因制作其睡眠相关的模型简便易行而得到广泛的应用,主要是采用各种物理化学方法制作失眠动物模型;转基因小鼠主要应用于药物干预睡眠及睡眠发生机制的研究;果蝇和斑马鱼多用于遗传学中睡眠的研究;猫用于睡眠研究的历史悠久,多用于体内试验,对内源性神经递质进行定量分析;猴睡眠结构与人相似,被广泛用于神经生物学,行为药理学等领域与睡眠的关系研究中.由于各种实验动物特殊的生理特征,在睡眠研究中多根据研究目的选择不同的实验动物.