Stem Cell

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    Protocols in Current Issue
    Mesenchymal Stromal Cells Derived from Bone Marrow and Adipose Tissue: Isolation, Culture, Characterization and Differentiation
    [Abstract] Since their discovery, mesenchymal stromal cells (MSCs) have received a lot of attention, mainly due to their self-renewal potential and multilineage differentiation capacity. For these reasons, MSCs are a useful tool in cell biology and regenerative medicine. In this article, we describe protocols to isolate MSCs from bone marrow (BM-MSCs) and ...
    Generation of BMEC Lines and in vitro BMEC-HSPC Co-culture Assays
    Authors:  Peipei Guo and Shahin Rafii, date: 11/05/2018, view: 3127, Q&A: 0
    [Abstract] Endothelial cells (ECs) sustain the self-renewal and regeneration of adult hematopoietic stem and progenitor cells (HSPCs) via deployment of EC-derived paracrine factors, termed as angiocrine factors. Generation of durable ex vivo vascular niche that maintains EC identity and preserves the angiocrine profile of organ of origin offers ...
    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: 4351, 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: 3991, 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 ...
    Cobblestone Area-forming Cell Assay of Mouse Bone Marrow Hematopoietic Stem Cells
    Authors:  Surya Amarachintha and Qishen Pang, date: 05/05/2018, view: 5469, 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 ...
    Adhesion Assay for Murine Bone Marrow Hematopoietic Stem Cells
    Authors:  Seymen Avci, Shiri Gur-Cohen, Francesca Avemaria and Tsvee Lapidot, date: 02/20/2017, view: 7324, Q&A: 0
    [Abstract] Hematopoietic stem cells (HSCs) are defined by their functional abilities to self-renew and to give rise to all mature blood and immune cell types throughout life. Most HSCs are retained in a non-motile quiescent state within a specialized protective microenvironment in the bone marrow (BM) termed the niche. HSCs are typically distinguished from ...
    VLA-4 Affinity Assay for Murine Bone Marrow-derived Hematopoietic Stem Cells
    Authors:  Francesca Avemaria, Shiri Gur-Cohen, Seymen Avci and Tsvee Lapidot, date: 02/20/2017, view: 5552, Q&A: 0
    [Abstract] Hematopoietic stem cells (HSCs) are defined by their functional ability to self-renew and to differentiate into all blood cell lineages. The majority of HSC reside in specific anatomical locations in the bone marrow (BM) microenvironment, in a quiescent non motile mode. Adhesion interactions between HSCs and their supporting BM microenvironment ...
    Isolation of Murine Adipose Tissue-derived Mesenchymal Stromal Cells (mASCs) and the Analysis of Their Proliferation in vitro
    Authors:  Per Anderson, Ana Belén Carrillo-Gálvez and Francisco Martín, date: 11/05/2015, view: 7899, Q&A: 0
    [Abstract] Mesenchymal stromal cells (MSCs) are non-hematopoietic, perivascular cells which support hematopoiesis and are thought to participate in tissue repair in vivo. MSCs can be isolated from various tissues and organs and are defined in vitro as plastic adherent cells expressing CD73, CD90, CD105 (human MSCs) or CD29, CD44, sca-1 ...
    PBMC-MSC Co-cultures for Induction of Treg Generation
    Authors:  Sara M. Melief, C. L. M. Schrama and Helene Roelofs, date: 01/20/2015, view: 11045, Q&A: 1
    [Abstract] To assess the capacity of multipotent stromal cells (MSC) to induce the generation of Tregs, transwell co-cultures were performed as well as cultures with MSC-conditioned medium (CM). In short, peripheral blood mononuclear cells (PBMC) were co-cultured with allogeneic MSC or CM for one week followed by one week of culture in the absence of MSC.
    Monocyte-MSC Co-cultures
    Authors:  Sara M. Melief, C. L. M. Schrama and Helene Roelofs, date: 01/20/2015, view: 8890, Q&A: 0
    [Abstract] To assess the effect of multipotent stromal cells (MSC) on monocytes, 3-day cultures were performed of freshly isolated monocytes in MSC-conditioned medium (CM). As a control condition, monocytes were stimulated with low dose macrophage colony-stimulating factor (M-CSF). Monocytes were isolated from peripheral blood mononuclear cell (PBMC) ...



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