Mouse multipotent mesenchymal stromal cell marker antibody panel

Manufactured by R&D Systems

The Mouse Multipotent Mesenchymal Stromal Cell Marker Antibody Panel is a collection of antibodies designed to identify and characterize mouse multipotent mesenchymal stromal cells. The panel includes a set of markers that are commonly used to phenotype and isolate this cell type.

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Lab products found in correlation

Adipogenesis assay kit, by Cayman Chemical (1 mentions) Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions) 100 mm culture dish, by BD (1 mentions) Wnt5a, by R&D Systems (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)

2 protocols using mouse multipotent mesenchymal stromal cell marker antibody panel

Isolation and Characterization of ADSCs

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ADSCs were prepared from C57BL/6 subcutaneous fat as described previously (Gondo et al., 2008 (link)) and cultured for 2 weeks in alpha-minimum essential medium containing 20% horse serum and 1% antibiotic antimycotic (Gibco) in a 5% CO₂ incubator at 37 °C. After four times passage of the culture, the adherent cells were used as ADSCs. For characterization of ADSCs, cell surface markers were analyzed by a flow cytometer using a Mouse Multipotent Mesenchymal Stromal Cell Marker Antibody Panel (R&D systems, Inc., MN). To test their capability for osteoblastic differentiation, ADSCs were cultured under a previously described condition (Gondo et al., 2004 (link)) and then stained with an anti-osteopontin antibody (R&D systems, Inc.). Induction of adipogenesis followed by Oil-Red O staining was performed using an Adipogenesis Assay Kit (Cayman Chemical Company, MI).

Tanaka T., Kojima D., Mera T., Matsumoto M., Yasunami Y, & Yanase T. (2018). Expansion of transplanted islets in mice by co-transplantation with adipose tissue-derived mesenchymal stem cells. Heliyon, 4(5), e00632.

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Isolation and Culture of Murine Mesenchymal Stem Cells

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Whole bone marrow cells (WBMCs) were isolated from wild-type (WT), Fanca^−/− and Fancc^−/− mice by gently flushing out of tibias and femurs using DPBS + 10% FBS. Cells obtained from two tibias and two femurs were pooled and plated in 100mm culture dish (BD Falcon, Tewksbury, MA) in 10 ml of MSCs media (Dulbecco’s Modified Eagle Medium (DMEM, Gibco, Gaithersburg, MD) supplemented with 15% fetal bovine serum (FBS, Gibco, Gaithersburg, MD) and 1% Penicillin-Streptomycin (Life Technologies # 15140-122 adapted and modified from previous report [27 (link)].
Plastic adherent cells were passaged 3 times followed by staining with fluorescently labelled antibodies for mesenchymal (CD29, CD44, CD73, SCA-1, CD106) and hematopoietic markers (CD45, CD11b) using the mouse multipotent mesenchymal stromal cell marker antibody panel (Cat # SC018; R&D systems; Minneapolis, MN) [28 –30 (link)]. We confirmed that at least 98% purity of MSCs was obtained with this culture method.
For Wnt5a treatment, MSCs at passages three were plated to obtain 95% confluence. Cells were pre-treated with different doses of Wnt5a (R&D System, Minneapolis, MN) for 16 hrs [31 (link)], followed by co-culture in fresh media with WT LSK (Lin⁻Sca1⁺c-kit⁺) cells.

Xu J., Li X., Cole A., Sherman Z, & Du W. (2018). Reduced Cdc42 activity compromises hematopoiesis-supportive function of Fanconi anemia mesenchymal stromal cells. Stem cells (Dayton, Ohio), 36(5), 785-795.

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