Protocols in Current Issue
    Skeletal Stem Cell Isolation from Cranial Suture Mesenchyme and Maintenance of Stemness in Culture
    Authors:  Takamitsu Maruyama, Hsiao-Man Ivy Yu and Wei Hsu, date: 03/05/2022, view: 601, Q&A: 0

    Skeletal stem cells residing in the suture mesenchyme are responsible for calvarial development, homeostatic maintenance, and injury-induced repair. These naïve cells exhibit long-term self-renewal, clonal expansion, and multipotency. They possess osteogenic abilities to regenerate bones in a cell-autonomous manner and can directly replace

    A Rapid Protocol for Direct Isolation of Osteoclast Lineage Cells from Mouse Bone Marrow

    Osteoclast lineage cells (OLCs), including osteoclast precursors (OCPs) and mature osteoclasts (MOCs), participate in bone remodeling and mediate pathologic bone loss. Thus, it is essential to obtain OLCs for exploring their molecular features in both physiological and pathological conditions in vivo. However, the conventional protocols for

    Isolation of CD31+ Bone Marrow Endothelial Cells (BMECs) from Mice
    Authors:  Alhaji Osman Smith, Seyram Yao Adzraku, Wen Ju, Jianlin Qiao, Kailin Xu and Lingyu Zeng, date: 11/20/2021, view: 1256, Q&A: 0

    In the bone marrow microenvironment, endothelial cells (ECs) play a pivotal role in regulating the production of both growth and inhibiting factors. They are held together by adherence molecules that interact with hematopoietic progenitor cells. The study of ECs in the hematopoietic stem cell niche is limited due to the lack of efficient protocols

    Metabolomic and Lipidomic Analysis of Bone Marrow Derived Macrophages
    Authors:  Gretchen L. Seim, Steven V. John and Jing Fan, date: 07/20/2020, view: 4646, Q&A: 0
    [Abstract] Macrophages are highly plastic immune cells that are capable of adopting a wide array of functional phenotypes in response to environmental stimuli. The changes in macrophage function are often supported and regulated by changes in cellular metabolism. Capturing a comprehensive picture of metabolism is vital for understanding the role of metabolic ...
    Analyzing the Quenchable Iron Pool in Murine Macrophages by Flow Cytometry
    Authors:  Michael Riedelberger and Karl Kuchler, date: 03/20/2020, view: 3695, Q&A: 0
    [Abstract] Tissue-resident macrophages are pivotal for a tightly-regulated iron metabolism at a cellular and systemic level, since subtle iron alterations increase the susceptibility for microbial infections or drive multiple diseases. However, research on cellular iron homeostasis in macrophages remains challenging due to the limited amount of available ...
    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 ...
    Bone-in-culture Array to Model Bone Metastasis in ex vivo Condition
    Authors:  Hai Wang and Xiang H.-F. Zhang, date: 01/20/2020, view: 2893, Q&A: 0
    [Abstract] Bone is the most frequently affected organ by metastases of breast cancer and prostate cancer. Our knowledge on bone metastasis is extremely limited due to the lack of potent and efficient experimental models. We developed the “Bone-In-Culture Array (BICA)” platform to model the bone colonization of cancer cells in ex vivo cultures. The ...
    Simultaneous Fluorescent Recordings of Extracellular ATP and Intracellular Calcium in Mammalian Cells
    Authors:  Nicholas Mikolajewicz and Svetlana V. Komarova, date: 05/20/2019, view: 4401, Q&A: 0
    [Abstract] Extracellular ATP is a potent signaling molecule that stimulates intracellular calcium responses through purinergic (P2) receptors in mammalian cells. While extracellular ATP and intracellular calcium can be measured separately, simultaneous monitoring can offer additional insights into P2 receptor physiology. This protocol takes advantage of the ...
    Cell Microencapsulation and Cryopreservation with Low Molecular Weight Hyaluronan and Dimethyl Sulfoxide
    Authors:  H. Gurruchaga, L. Saenz del Burgo, G. Orive, R. M. Hernandez, J. Ciriza and J. L. Pedraz, date: 02/20/2019, view: 4205, Q&A: 0
    [Abstract] Cryopreservation is commonly used for the storage of cells, tissues, organs or 3D cell-based products using ultra-low temperatures, which involves the immersion in liquid nitrogen for their long-term preservation. The cryopreservation of several microencapsulated cells is usually performed by the slow freezing with the dimethyl sulfoxide (DMSO) as ...
    Calvarial Bone Implantation and in vivo Imaging of Tumor Cells in Mice
    Authors:  Kyoko Hashimoto, Shingo Sato, Hiroki Ochi, Shu Takeda and Mitsuru Futakuchi, date: 02/05/2019, view: 5580, Q&A: 0
    [Abstract] Bone is one of common metastasis sites for many types of cancer. In bone metastatic microenvironment, tumor-bone interactions play a significant role in the regulation of osteolytic or osteoblastic bone metastasis. In order to investigate the direct interaction between tumor cells and bone tissue, it is essential to generate appropriate animal ...

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