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Stem Cell

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    Protocols in Current Issue
    Small Molecule-Based Retinal Differentiation of Human Embryonic Stem Cells and Induced Pluripotent Stem Cells
    Authors:  Jie Zhu and Deepak A. Lamba, date: 06/20/2018, view: 659, Q&A: 0
    [Abstract] Retinal degeneration leads to loss of light-sensing photoreceptors eventually resulting in vision impairment and impose a heavy burden on both patients and the society. Currently available treatment options are very limited and mainly palliative. Ever since the discovery of human pluripotent stem cell technologies, cell replacement therapy has ...
    Expansion of Airway Basal Cells and Generation of Polarized Epithelium
    Authors:  Hannah Levardon, Lael M. Yonker, Bryan P. Hurley and Hongmei Mou, date: 06/05/2018, view: 602, Q&A: 0
    [Abstract] Airway basal stem cells are the progenitor cells within the airway that exhibit the capacity to self-renew and give rise to multiple types of differentiated airway epithelial cells. This stem cell-derived epithelium displays organized architecture with functional attributes of the airway mucosa. A protocol has been developed to culture and expand ...
    High Dimensional Functionomic Analysis of Human Hematopoietic Stem and Progenitor Cells at a Single Cell Level
    Authors:  Thomas Luh, Kimberly Lucero, Wenji Ma, Jaeyop Lee, Yu Jerry Zhou, Yufeng Shen and Kang Liu, date: 05/20/2018, view: 815, Q&A: 0
    [Abstract] The ability to conduct investigation of cellular transcription, signaling, and function at the single-cell level has opened opportunities to examine heterogeneous populations at unprecedented resolutions. Although methods have been developed to evaluate high-dimensional transcriptomic and proteomic data (relating to cellular mRNA and protein), ...
    Osteoblast Sorting and Intracellular Staining of CXCL12
    Authors:  Weihuan Wang, Gurnoor Majhail, Cui Lui and Lan Zhou, date: 05/20/2018, view: 638, Q&A: 0
    [Abstract] Osteoblasts are bone marrow endosteum-lining niche cells playing important roles in the regulation of hematopoietic stem cells by secreting factors and cell adhesion molecules. Characterization of primary osteoblasts has been achieved through culture of outgrowth of collagenase treated bone. Immunophenotyping and flow-based analysis of long bone ...
    A Method to Injure, Dissect and Image Indirect Flight Muscle of Drosophila
    Authors:  Kunal Chakraborty, K. VijayRaghavan and Rajesh Gunage, date: 05/20/2018, view: 1147, Q&A: 0
    [Abstract] Inducing an injury specifically to Drosophila flight muscles is a difficult task, owing to the small size of the muscles and the presence of the cuticle. The protocol described below provides an easy and reproducible method to induce injury in the Drosophila flight muscles.
    Murine Hair Follicle Derived Stem Cell Transplantation onto the Cornea Using a Fibrin Carrier
    [Abstract] The goal of this protocol is to establish a procedure for cultivating stem cells on a fibrin carrier to allow for eventual transplantation to the eye. The ability to transfer stem cells to a patient is critical for treatment for a variety of disorders and wound repair. We took hair follicle stem cells from the vibrissae of transgenic mice ...
    Hair Follicle Stem Cell Isolation and Expansion
    [Abstract] Stem cells are widely used for numerous clinical applications including limbal stem cell deficiency. Stem cell derived from the bulge region of the hair follicle have the ability to differentiate into a variety of cell types including interfollicular epidermis, hair follicle structures, sebaceous glands and corneal epithelial cells when provided ...
    Cobblestone Area-forming Cell Assay of Mouse Bone Marrow Hematopoietic Stem Cells
    Authors:  Surya Amarachintha and Qishen Pang, date: 05/05/2018, view: 811, Q&A: 0
    [Abstract] Bone Marrow Hematopoietic Stem Cells (HSCs) require bone marrow microenvironment for their maintenance and proliferation. Culture of Bone Marrow Mesenchymal Stromal Cells (MSCs) provides appropriate environmental signals for HSCs survival in vitro. Here, we provide a detailed protocol that describes culture conditions for MSCs, flow ...
    Generating Loss-of-function iPSC Lines with Combined CRISPR Indel Formation and Reprogramming from Human Fibroblasts
    Authors:  Andrew M. Tidball, Preethi Swaminathan, Louis T. Dang and Jack M. Parent, date: 04/05/2018, view: 1577, Q&A: 0
    [Abstract] For both disease and basic science research, loss-of-function (LOF) mutations are vitally important. Herein, we provide a simple stream-lined protocol for generating LOF iPSC lines that circumvents the technical challenges of traditional gene-editing and cloning of established iPSC lines by combining the introduction of the CRISPR vector ...
    Primary Cultures from Human GH-secreting or Clinically Non-functioning Pituitary Adenomas
    Authors:  Roberto Würth, Alessandra Pattarozzi, Federica Barbieri and Tullio Florio, date: 04/05/2018, view: 810, Q&A: 0
    [Abstract] Pituitary adenomas are among the more frequent intracranial tumors usually treated with both surgical and pharmacological–based on somatostatin and dopamine agonists–approaches. Although mostly benign tumors, the occurrence of invasive behaviors is often detected resulting in poorer prognosis. The use of primary cultures from human pituitary ...