Biliary atresia (BA) is a common cause of pediatric end-stage liver disease. While its etiology is not yet clear, evidence has suggested that BA results from interactions between genetic ...susceptibility and environmental factors. Disease relevant human cellular models of BA will facilitate identification of both genetic and environmental factors that are important for disease prevention and treatment. Here we report the generation of a human induced pluripotent stem cell line from a BA patient using episomal vectors. Patient-specific BA iPSC lines provide valuable tools for disease mechanism study and drug development.
Pluripotent human embryonic stem (hES) cells can differentiate into various cell types derived from the three embryonic germ layers and extraembryonic tissues such as trophoblasts. The mechanisms ...governing lineage choices of hES cells are largely unknown. Here, we report that we established two independent hES cell clones lacking a group of cell surface molecules, glycosyl-phosphatidyl-inositol-anchored proteins (GPI-APs). The GPI-AP deficiency in these two hES clones is due to the deficiency in the gene expression of PIG-A (phosphatidyl-inositol-glycan class A), which is required for the first step of GPI synthesis. GPI-AP-deficient hES cells were capable of forming embryoid bodies and initiating cell differentiation into the three embryonic germ layers. However, GPI-AP-deficient hES cells failed to form trophoblasts after differentiation induction by embryoid body formation or by adding exogenous BMP4. The defect in trophoblast formation was due to the lack of GPI-anchored BMP coreceptors, resulting in the impairment of full BMP4 signaling activation in the GPI-AP-deficient hES cells. These data reveal that GPI-AP-enhanced full activation of BMP signaling is required for human trophoblast formation.
Molecular Imaging and Stem Cell Research Jang, Yoon-Young; Ye, Zhaohui; Cheng, Linzhao
Molecular Imaging,
03/2011, Letnik:
10, Številka:
2
Book Review, Journal Article
Recenzirano
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During the last decade, there has been enormous progress in understanding both multipotent stem cells such as hematopoietic stem cells and pluripotent stem cells such as embryonic stem cells and ...induced pluripotent stem cells. However, it has been challenging to study developmental potentials of these stem cells because they reside in complex cellular environments and aspects of their distribution, migration, engraftment, survival, proliferation, and differentiation often could not be sufficiently elucidated based on limited snapshot images of location or environment or molecular markers. Therefore, reliable imaging methods to monitor or track the fate of the stem cells are highly desirable. Both short-term and more permanent monitoring of stem cells in cultures and in live organisms have benefited from recently developed imaging approaches that are designed to investigate cell behavior and function. Confocal and multiphoton microscopy, time-lapse imaging technology, and series of noninvasive imaging technologies enable us to investigate cell behavior in the context of a live organism. In turn, the knowledge gained has brought our understanding of stem cell biology to a new level. In this review, we discuss the application of current imaging modalities for research of hematopoietic stem cells and pluripotent stem cells and the challenges ahead.
Stem cells are characterized by the properties of self-renewal and the ability to differentiate into multiple cell types, and thus maintain tissue homeostasis. Reactive oxygen species (ROS) are a ...natural byproduct of aerobic metabolism and have roles in cell signaling. Regulation of ROS has a vital role in maintaining "stemness" and differentiation of the stem cells, as well as in progression of stem-cell-associated diseases.
As of late, much research has been done on the adverse effects of ROS in stem cells. However, recently it has become apparent that in some cases redox status of the stem cell does have a role in maintaining its identity as such. Both pluripotent and multipotent stem cell types have been reported to possess enzymatic and nonenzymatic mechanisms for detoxification of ROS and to correct oxidative damage to the genome as well as the proteome.
Although context dependent and somewhat varied among different stem cell types, the correlation seems to exist between antioxidant defense level and stem cell fate change (i.e., proliferation, differentiation, and death). Changes in stem cell redox regulation may affect the pathogenesis of various human diseases.
Dissecting the defined roles of ROS in distinct stem cell types will greatly enhance their basic and translational applications. Here, we discuss the various roles of ROS in adult, embryonic, and induced pluripotent stem cells.
In addition to being an attractive source for cell replacement therapy, human induced pluripotent stem cells (iPSCs) also have great potential for disease modeling and drug development. During the ...recent several years, cell reprogramming technologies have evolved to generate virus-free and integration-free human iPSCs from easily accessible sources such as patient skin fibroblasts and peripheral blood samples. Hematopoietic cells from umbilical cord blood banks and Epstein Barr virus (EBV) immortalized B lymphocyte repositories represent alternative sources for human genetic materials of diverse backgrounds. Ability to reprogram these banked blood cells to pluripotency and differentiate them into a variety of specialized and functional cell types provides valuable tools for studying underlying mechanisms of a broad range of diseases including rare inherited disorders. Here we describe the recent advances in generating disease specific human iPSCs from these different types of hematopoietic cells and their potential applications in disease modeling and regenerative medicine.
Hematopoietic stem cells (HSCs) represent an important target for the treatment of various blood disorders. As the source of critical cells within the immune system, genetic modification of HSCs can ...also be used to modulate immune responses. The effectiveness of HSC-mediated gene therapy largely depends on efficient gene delivery into long-term repopulating progenitors and targeted transgene expression in an appropriate progeny of the transduced pluripotent HSCs. Self-inactivating (SIN) lentiviral vectors have been demonstrated to be capable of transducing mitotically inactive cells, including HSCs, and accommodating a nonviral promoter to control the transgene expression in transduced cells. In this study, we constructed 2 SIN lentiviral vectors, EF.GFP and DR.GFP, to express the green fluorescent protein (GFP) gene controlled solely by the promoter of either a housekeeping gene EF-1α or the human HLA-DRα gene, which is selectively expressed in antigen-presenting cells (APCs). We demonstrated that both vectors efficiently transduced human pluripotent CD34+cells capable of engrafting nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice. When the EF.GFP vector was used, constitutive high-level GFP expression was obtained in all the human HSC progeny detectable in NOD/SCID mice and in subsequent in vitro differentiation assays, indicating that engrafting human HSCs have been transduced. In contrast, the DR.GFP vector mediated transgene expression specifically in human HLA-DR+ cells and highly in differentiated dendritic cells (DCs), which are critical in regulating immunity. Furthermore, human DCs derived from transduced and engrafted human cells potently stimulated allogeneic T-cell proliferation. This study demonstrated successful targeting of transgene expression to APCs/DCs after stable gene transduction of pluripotent HSCs.
To mitigate the sudden increase in the production of waste engineering slurry, predominantly composed of Kaolinite, this study investigated the flocculation and dewatering of Kaolin slurry treated ...with single- and dual-polymer flocculants. The influence of the flocculant type and dosage, under single- and dual-dose conditions, on slurry's sedimentation and the filtration characteristics, were thoroughly discussed. The results reveal that the adsorption bridging of the polymeric flocculant, resulting from hydrogen bonds, exerts a more significant effect than electrical neutralization on forming a large floc. Under single-dose conditions, nonionic polyacrylamides (NPAMs) with the strongest adsorption bridging leads to biggest flocs and the maximum settling rate of 21.55 mm/s. Under the dual-dose conditions of polymeric aluminium chloride (PAC) and PAM, the size of the slurry's floc decreases with an increase in PAC dosage. Nevertheless, the filtration performance of the slurry improves, with the lowest SRF value of the flocculated slurry being 1.58 × 1011 m/kg as 3‰ PAC and 3‰ NPAM is dosed. The improvement is explained by the micro-pore distribution of sludge. According to Mercury intrusion porosimetry (MIP) test, the slurry treated with the optimal dosage of dual-polymer flocculant exhibits the greatest sludge pore size and connected porosity (with a maximum value of 20.99%). Furthermore, the study discusses and compares the flocculation mechanism of single- and dual-polymer flocculants. The obtained results provide guidance for selecting appropriate flocculants for dewatering inorganic slurries, using different dewatering methods, such as gravitational thickening or filter pressing.
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•Adsorption bridging effect decides flocculation and dewatering of Kaolin slurry.•Multiple flocculation effects of dual flocculants reduce the Kaolin's settling rate.•Dual flocculants can improve the slurry's filtration performance.•Filtration depends on the sludge's pore size distribution and connectivity.
Independent component analysis is intended to recover the mutually independent components from their linear mixtures. This technique has been widely used in many fields, such as data analysis, signal ...processing, and machine learning. To alleviate the dependency on prior knowledge concerning unknown sources, many nonparametric methods have been proposed. In this paper, we present a novel boosting-based algorithm for independent component analysis. Our algorithm consists of maximizing likelihood estimation via boosting and seeking unmixing matrix by the fixed-point method. A variety of experiments validate its performance compared with many of the presently known algorithms.
The last decade has seen many exciting technological breakthroughs that greatly expanded the toolboxes for biological and biomedical research, yet few have had more impact than induced pluripotent ...stem cells and modern-day genome editing. These technologies are providing unprecedented opportunities to improve physiological relevance of experimental models, further our understanding of developmental processes, and develop novel therapies. One of the research areas that benefit greatly from these technological advances is the three-dimensional human organoid culture systems that resemble human tissues morphologically and physiologically. Here we summarize the development of human pluripotent stem cells and their differentiation through organoid formation. We further discuss how genetic modifications, genome editing in particular, were applied to answer basic biological and biomedical questions using organoid cultures of both somatic and pluripotent stem cell origins. Finally, we discuss the potential challenges of applying human pluripotent stem cell and organoid technologies for safety and efficiency evaluation of emerging genome editing tools.
Nonparametric maximum likelihood estimation is intended to infer the unknown density distribution while making as few assumptions as possible. To alleviate the over parameterization in nonparametric ...data fitting, smoothing assumptions are usually merged into the estimation. In this letter a novel boosting-based method is introduced to the nonparametric estimation in univariate cases. We deduce the boosting algorithm by the second-order approximation of nonparametric log-likelihood. Gaussian kernel and smooth spline are chosen as weak learners in boosting to satisfy the smoothing assumptions. Simulations and real data experiments demonstrate the efficacy of the proposed approach.
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