NDLI: Cobblestone Area-forming Cell Assay of Mouse Bone Marrow Hematopoietic Stem Cells.

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Cobblestone Area-forming Cell Assay of Mouse Bone Marrow Hematopoietic Stem Cells.

Content Provider	Europe PMC
Author	Amarachintha, Surya Pang, Qishen
Copyright Year	2018
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 cytometric isolation of HSCs from mouse bone marrow, and perform co-culture of MSCs and HSCs known as Cobblestone area-forming cell (CAFC) assay. Altogether, CAFC assays can be used as a high-throughput in vitro screening model where efforts are made to understand and develop therapies for complex bone marrow diseases. This protocol needs 3 to 4 weeks starting from culturing MSCs, isolating LSK cells (HSCs), and to performing limited dilution CAFC assay.
Related Links	https://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC8275301&blobtype=pdf
Volume Number	8
PubMed Central reference number	PMC8275301
Issue Number	9
PubMed reference number	34286035
Journal	Bio-protocol [Bio Protoc]
e-ISSN	23318325
DOI	10.21769/bioprotoc.2824
Language	English
Publisher	Bio-Protocol
Publisher Date	2018-05-05
Publisher Place	1075 Lorne Way, Sunnyvale, CA 94087, USA
Access Restriction	Open
Rights License	Copyright © 2018 The Authors; exclusive licensee Bio-protocol LLC.
Subject Keyword	Mesenchymal Stromal Cells Hematopoietic Stem Cells Co-culture assay Cobblestone area-forming cell assay
Content Type	Text
Resource Type	Article
Subject	Immunology and Microbiology Neuroscience Plant Science Biochemistry, Genetics and Molecular Biology

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