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细胞功能测定

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Isolation and Purification of Murine Male Germ Cells

Seminiferous tubules of the male testis contain somatic cells (Sertoli and Leydig cells) and germ cells at different stages of spermatogenesis (spermatogonia, spermatocytes and spermatids). Germ cells at different stages of differentiation migrate toward the central lumen via c ...

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Isolation of Human Male Germ-Line Stem Cells Using Enzymatic Digestion and Magnetic-Activated Cell Sorting

Mammalian spermatogenesis is a process whereby male germ-line stem cells (spermatogonial stem cells) divide and differentiate into sperm. Although a great deal of progress has been made in the isolation and characterization of spermatogonial stem cells (SSCs) in rodents, little is kn ...

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Isolation of Undifferentiated and Early Differentiating Type A Spermatogonia from Pou5f1-GFP Reporter Mice

Limited understanding of the mechanisms underlying self-renewal and differentiation of spermatogonial stem cells hampers our ability to develop new therapeutic and contraceptive approaches. Mouse models of spermatogonial stem cell development are key to developing new in ...

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Neonatal Testicular Gonocytes Isolation and Processing for Immunocytochemical Analysis

In recent years, an increasing interest has emerged at understanding how spermatogonial stem cells (SSCs) arise from their precursor cells, the gonocytes. The identification of factors acting directly on gonocytes rather than on the surrounding somatic cells and the study of genes and si ...

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Isolation of Fetal Gonads from Embryos of Timed-Pregnant Mice for Morphological and Molecular Studies

Gonadal sex differentiation is an important developmental process, in which a bipotential primordial gonad undergoes two distinct pathways, i.e., testicular and ovarian differentiation, dependent on its genetic sex. Techniques of isolating fetal gonads at various developme ...

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Spermatogenesis in Cryptorchidism

Cryptorchidism or undescended testis is the most frequent congenital abnormality in newborn boys. The process of testicular descent to the scrotum is controlled by hormones produced in Leydig cells, insulin-like3, and androgens. Variation in genetic and environmental factors mi ...

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Methylation Profiling Using Methylated DNA Immunoprecipitation and Tiling Array Hybridization

DNA methylation is an important epigenetic modification that regulates development and plays a role in the pathophysiology of many diseases. It is dynamically changed during germline development. Methylated DNA immunoprecipitation (MeDIP) is an efficient, cost-effective m ...

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Closed Ampoule Isothermal Microcalorimetry for Continuous Real-Time Detection and Evaluation of Cultured Mammalian Cell Activity and Responses

Closed ampoule isothermal microcalorimetry (IMC) is a simple, powerful, nondestructive, and convenient technique that allows continuous, real-time detection and evaluation of cultured cell activity and responses. At a selected set temperature, IMC measures the heat flow betwe ...

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Cell Viability Assays: Introduction

The measurement of cell viability plays a fundamental role in all forms of cell culture. Sometimes it is the main purpose of the experiment, such as in toxicity assays. Alternatively, cell viability can be used to �correlate cell behaviour to cell number, providing a more accurate picture of, for exa ...

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Raman Micro-Spectroscopy as a Non-invasive Cell Viability Test

The number of techniques to identify, quantify and characterise cell death is rapidly increasing as more is known about the complex mechanisms underlying this process. However, most of these techniques are invasive and require preparation steps such as cell fixation, staining or protein ...

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Noninvasive Bioluminescent Quantification of Viable Stem Cells in Engineered Constructs

Bioluminescence from murine stem cells tagged with the luciferase gene reporter and distributed within three-dimensional scaffolds of two different materials is quantified in vitro and in vivo. The luminescence emitted from cells adhering to the scaffolds tested is monitored non ...

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Exploiting the Liberation of Zn2+ to Measure Cell Viability

Zn2+ ions are a critical component of cellular machinery. The ion is required for the function of many cell components crucial to survival, such as transcription factors, protein synthetic machinery, metabolic enzymes, hormone packaging, among other roles. In stark contrast to the cells’ n ...

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Measuring Glutamate Receptor Activation-Induced Apoptotic Cell Death in Ischemic Rat Retina Using the TUNEL Assay

Glutamate receptor activation-mediated excitotoxicity has been hypothesized to cause cell death in both acute and chronic neurodegenerative diseases including glaucoma. Although the precise mechanisms of ischemia-induced neuronal death are unknown, glutamate excit ...

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Viability Assessment of Osteocytes Using Histological Lactate Dehydrogenase Activity Staining on Human Cancellous Bone Sections

The assessment of viable osteocytes within bone tissue is of crucial importance. Osteocytes are the most abundant cells in bone. Due to their interconnectivity in the bone matrix they are hypothesised to play an important role in the maintenance of the extracellular matrix of bone. The death of o ...

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Confocal Imaging Protocols for Live/Dead Staining in Three-Dimensional Carriers

In tissue engineering, a variety of methods are commonly used to evaluate survival of cells inside tissues or three-dimensional (3D) carriers. Among these methods confocal laser scanning microscopy opened accessibility of 3D tissue using live cell imaging into the tissue or 3D scaffold ...

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Fluorescein Diacetate for Determination of Cell Viability in 3D FibroblastCollagenGAG Constructs

Quantification of cell viability and distribution within engineered tissues currently relies on representative histology, phenotypic assays, and destructive assays of viability. To evaluate uniformity of cell density throughout 3D collagen scaffolds prior to in vivo use, a no ...

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Cytotoxicity Testing: Measuring Viable Cells, Dead Cells, and Detecting Mechanism of Cell Death

Testing the effects of compounds on the viability of cells grown in culture is widely used as a predictor of potential toxic effects in whole animals. Among the several alternative assays available, measuring the levels of ATP is the most sensitive, reliable, and convenient method for monitori ...

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Multiplexing Cell Viability Assays

Today, obtaining mechanistic insights into biological, toxicological, and pathological processes is of upmost importance. Researchers aim to obtain as many as possible data from one cell sample to understand the biological processes under study. Multiplexing, which is the ability ...

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Rapid Quantification of Cell Viability and Apoptosis in B-Cell Lymphoma Cultures Using Cyanine SYTO Probes

The gross majority of classical apoptotic hallmarks can be rapidly examined by multiparameter flow cytometry. As a result, cytometry became a technology of choice in diverse studies of cellular demise. In this context, a novel class of substituted unsymmetrical cyanine SYTO probes has re ...

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Measurement of Caspase Activity: From Cell Populations to Individual Cells

Caspases are critical regulators of the apoptotic program, responsible for the harmonic dismantling of the cell. Cell death can occur by way of different options (necroptosis, necrosis, extreme autophagy) but once caspases are fully engaged it will take the apoptotic route. Hence, in gene ...

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