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细胞功能测定

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Purification of Adipose Tissue Mesenchymal Stem Cells and Differentiation Toward Hepatic-Like Cells

There is a great interest in the development of functional hepatocytes in vitro from different types of stem cells. Multipotential mesenchymal stem cells (MSC) compose a great source for stem cell based therapy, especially, because they can be obtain from patients own tissues, sidesteppi ...

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SAMe and HuR in Liver Physiology: Usefulness of Stem Cells in Hepatic Differentiation Research

S-Adenosylmethionine, abbreviated as SAM, SAMe or AdoMet, is the principal methyl group donor in the mammalian cell and the first step metabolite of the methionine cycle, being synthesized by MAT (methionine adenosyltransferase) from methionine and ATP. About 60 years after its identi ...

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Tet-On System Toward Hepatic Differentiation of Human Mesenchymal Stem Cells by Hepatocyte Nuclear Factor

“Tet-On” system requires two DNA constructs: the first one is a transcription regulatory unit, rtTA and the second construct is the responsive element Escherichia coli sequences (tetO) linked to Pcmv driven target gene. In the absence of inducing agent doxycycline (Dox), a tetracycline der ...

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Efficient Hepatic Differentiation from Human iPS Cells by Gene Transfer

Establishment of protocols for the differentiation of hepatic cells from human embryonic stem (ES) and induced pluripotent stem (iPS) cells could contribute to regenerative cell therapies or drug discovery and development. However, the differentiation efficiency of endoderm ...

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Isolation of Hepatic Progenitor Cells from the Galactosamine-Treated Rat Liver

Oval cells and small hepatocytes (SHs) are well known as hepatic stem/progenitor cells. However, the relationship between the oval cells and SHs in liver regeneration is not well understood. To resolve this issue, we established a technique to selectively separate oval cells and SHs. In the inj ...

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Isolation and Purification Method of Mouse Fetal Hepatoblasts

During development, liver precursors constitute a valuable source of pluripotent stem cells that present the ability to differentiate into both a hepatic and biliary lineage. In the present chapter, we report an experimental procedure developed by our group to isolate mouse fetal hepa ...

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Identification and Isolation of Adult Liver Stem/Progenitor Cells

Hepatoblasts are considered to be liver stem/progenitor cells in the fetus because they propagate and differentiate into two types of liver epithelial cells, hepatocytes and cholangiocytes. In adults, oval cells that emerge in severely injured liver are considered facultative hep ...

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Clinical Uses of Liver Stem Cells

Liver transplantation offers a definitive cure for many liver and metabolic diseases. However, the complex invasive procedure and paucity of donor liver graft organs limit its clinical applicability. Liver stem cells provide a potentially limitless source of cells that would be usef ...

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Purification and Culture of Fetal Mouse Hepatoblasts that Are Precursors of Mature Hepatocytes and Biliary Epithelial Cells

To investigate cell–cell interactions during mammalian liver development, it is essential to separate hepatoblasts (fetal liver progenitor cells) from nonparenchymal cells, including stellate cells, endothelial cells, and hemopoietic cells. Various factors, which may be p ...

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Gene Expression in Mouse Oocytes by RNA-Seq

Second-generation sequencers such as the Illumina GAIIx make possible the study of all transcribed loci in a genome across an almost endless dynamic range. Although typical protocols call for starting from at least 1 μg of total RNA, this is not possible when studying small tissues or rare cell typ ...

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DNA Methylation Analysis of Germ Cells by Using Bisulfite-Based Sequencing Methods

Dynamic changes in DNA methylation at the gene-specific and genome-wide level occur during mammalian germ-cell development. However, the details of how the methylation profiles change remain largely unknown. Bisulfite sequencing analysis is a powerful technique to determine the ...

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Investigating the Origins of Somatic Cell Populations in the Perinatal Mouse Ovaries Using Genetic Lineage Tracing and Immunohistochemistry

Genetic lineage tracing (or fate mapping) techniques are designed to permanently label progenitor cells of target tissues, thereby allowing delineation of the progenies of labeled cells during organogenesis. This technology has been widely used in the study of cell migration and lin ...

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Experimental Approaches to the Study of Human Primordial Germ Cells

The survival, proliferation, and differentiation of primordial germ cells in the mammalian embryo is regulated by a complex cocktail of growth factors and interactions with surrounding somatic cells, which together form a microenvironment known as the germ cell niche. Extensive ins ...

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In Vitro Maturation (IVM) of Porcine Oocytes

Oocyte maturation is a critical component of in vitro embryo production. If not carried out in a precise manner under optimal conditions, subsequent fertilization and embryo development will be compromised. Here, we describe collection and in vitro maturation procedures in swine that m ...

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Microspread Ovarian Cell Preparations for the Analysis of Meiotic Prophase Progression in Oocytes with Improved Recovery by Cytospin Centrifugation

Female fertility is critically influenced by two events affecting oocytes during meiotic prophase progression: meiotic recombination between homologous chromosomes; and a major oocyte loss. It is technically challenging to examine these events, which take place in fetal and neo ...

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In Vitro Culture of Fetal Ovaries: A Model to Study Factors Regulating Early Follicular Development

Follicular development commences with the formation of primordial follicles, which begins with the differentiation of pluripotent ovarian somatic cells into early granulosa cells and their apposition to the oocytes in the egg nest. The process of primordial follicle morphogene ...

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Identification of Novel Long Noncoding RNA Transcripts in Male Germ Cells

Emerging evidence from these studies suggested that the male germ cell transcriptome is more complex than previously envisioned. In addition to protein-coding genes, the transcriptome also encodes a significant number of nonprotein-coding transcripts. These noncoding (nc) RN ...

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Biochemical Characterization of a Testis-Predominant Isoform of N-Alpha Acetyltransferase

N-alpha protein acetylation, catalyzed by N-alpha acetyltransferase complex, is a common protein �modification process in eukaryotic cells. Despite its widespread occurrence, the biological significance of this modification process is still unclear. We recently discove ...

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Revealing the Transcriptome Landscape of Mouse Spermatogonial Cells by Tiling Microarray

In the past decade, the advent of microarray technologies has allowed functional genomic studies of male germ cell development, resulting in the identification of genes governing various processes. A major limitation with conventional gene expression microarray is that results o ...

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Preparation of Enriched Mouse Syncitia-Free Pachytene Spermatocyte Cell Suspensions

Spermatogenesis comprises a complex succession of steps of mitosis, meiosis, and differentiation, starting with the commitment of diploid spermatogonial stem cells to differentiate and ending with the formation of haploid spermatozoa. Rodent models have been routinely used to s ...

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