Human msc analysis kit

Manufactured by BD

Sourced in United States

The Human MSC Analysis Kit is a laboratory equipment product that enables the analysis of human mesenchymal stem cells (MSCs). It provides the core functionality to facilitate the study and characterization of these cell types.

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Lab products found in correlation

Facscanto 2, by BD (15 mentions) Accutase, by BD (6 mentions) Accutase cell detachment solution, by BD (6 mentions) Stain buffer, by BD (5 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) Facscanto, by BD (4 mentions) Facsdiva software, by BD (4 mentions) Facscalibur flow cytometer, by BD (3 mentions) Facscalibur, by BD (3 mentions) Facsaria 2, by BD (3 mentions) Facsaria 2 cell sorter, by BD (2 mentions) Accutase, by Merck Group (2 mentions) Lsrfortessa flow cytometer, by BD (2 mentions) Tryple, by Thermo Fisher Scientific (2 mentions) Trypsin, by Thermo Fisher Scientific (2 mentions) Human msc phenotyping kit, by Miltenyi Biotec (2 mentions) Percp cy5, by BD (2 mentions) Cell wash buffer, by BD (2 mentions) Oil red o, by Merck Group (2 mentions) Facscalibur 2 cytometer, by BD (2 mentions)

Spelling variants of this product

BD Stemflow™ Human MSC Analysis Kit (24 citations) MSC Analysis Kit (7 citations) Human MSCs Analysis Kit (3 citations)

101 protocols using human msc analysis kit

Phenotypic Characterization of MSCs

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To test the surface markers of these two types of MSCs, 500 μL single cell suspension prepared from P4-generation hADSCs and hUCMSCs with 0.25% trypsin was mixed with CD90-FITC, CD44-PE, CD73-APC and CD105-PerCP-P5.5 antibodies provided in the human MSC analysis kit (562245; BD Biosciences, Franklin Lakes, USA) and incubated on ice for 30 min. Then the mixture was centrifuged at 200
g for 10 min, and the cell pellet was washed three times with PBS and detected by flow cytometry with a FACSVerse flow cytometer (BD Biosciences).

Jin Y., Shi R., Qi T., Li Q., Chen C., Gao S., Gao F., Yang D., Sun G., Xu J., Fu Q., Xu J, & Zhang X. (2023). Adipose-derived stem cells show hepatic differentiation potential and therapeutic effect in rats with acute liver failure: Adipose-derived stem cells for acute liver failure. Acta Biochimica et Biophysica Sinica, 55(4), 601-612.

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Characterization of Human MSCs

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BMSCs were purchased from Chinese Academy of Sciences and kept in NutriStem® XF Medium. Surface antigens of MSCs were characterized using a CytoFLEX Flow Cytometer (Beckman, Beijing) and the Human MSC Analysis Kit (BD Biosciences).

Chen C., Zhu F., Liu F., Yao Y., Ma Z, & Luo S. (2023). Human Bone Marrow Mesenchymal Stem Cells-Derived Exosomal miRNA-21-5p Inhibits Lidocaine-Induced Apoptosis in SH-SY5Y Neuroblastoma Cells. Iranian Journal of Public Health, 52(4), 756-765.

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Phenotypic Characterization of HGMSCs

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Phenotypic characterization was carried out according to ISCT guidelines. In brief, approximately 6 × 10⁶ HGMSCs were incubated with a Human MSC Analysis kit (BD) containing preconjugated and pretitrated cocktails with defined positive and negative expression markers along with the corresponding isotype controls.^{8 (link)} It was subjected to analysis using a BD FACSCalibur flow cytometer.

Rao S.R., Subbarayan R., Dinesh M.G., Arumugam G, & Raja S.T. (2016). Differentiation of human gingival mesenchymal stem cells into neuronal lineages in 3D bioconjugated injectable protein hydrogel construct for the management of neuronal disorder. Experimental & Molecular Medicine, 48(2), e209-.

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Expansion and Characterization of Human Mesenchymal Stem Cells

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Human mesenchymal stem cells were purchased from ATCC (catalog: PCS-500-012) and grown per manufacturer recommendations in basal medium (mesenchymal stem cell basal medium for adipose, umbilical, and bone marrow-derived MSCs, catalog: PCS-500-030) with supplementation (mesenchymal stem cell growth kit for bone marrow-derived MSCs, catalog: PCS-500-041) also purchased from ATCC. Cells were maintained at 37°C in an atmosphere of 5% carbon dioxide. Using the human MSC analysis kit (BD Biosciences), cells were found to be conforming to the definition of stemness as outlined by the International Society for Cellular Therapy (25 (link)). After two passages using Accutase (STEM CELL Technologies), cells were verified to be mycoplasma free, and stocks were made in 10% DMSO at a concentration of about 1 × 10⁶ cells per vial. HUVECs, GH354 cells, and mDCT cells were also purchased from ATCC and grown as per manufacturer’s recommendations in their respective culture media. Antibiotics were not used in any step of the process.

Suresh R.V., Deng B., Gebremicale Y., Roche K., Miura K, & Long C. (2023). Mesenchymal stem cells of the bone marrow raise infectivity of Plasmodium falciparum gametocytes. mBio, 14(6), e02232-23.

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Characterization of Human Mesenchymal Stem Cells

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HucMSCs were provided by Stem Cell Bank, Chinese Academy of Sciences and cultured in serum-free MSC NutriStem® XF Medium (Biological Industries, Beit HaEmek, Israel). When the cells grew to 80–90% confluence, they were digested with 0.25% trypsin containing 0.01% EDTA. The cells were resuspended in PBS and adjusted to 1 × 10⁶ cells/ml. The mouse antihuman CD34, CD45, CD44, and CD105 were added and incubated at 4°C in dark for 15–30 min. Surface antigens of MSCs were characterized by using a Beckman CytoFLEX flow cytometer (Beckman Coulter Life Sciences, Tokyo, Japan) and the Human MSC Analysis Kit (BD Biosciences, San Jose, CA, United States).

Zhong Z., Tian Y., Luo X., Zou J., Wu L, & Tian J. (2021). Extracellular Vesicles Derived From Human Umbilical Cord Mesenchymal Stem Cells Protect Against DOX-Induced Heart Failure Through the miR-100-5p/NOX4 Pathway. Frontiers in Bioengineering and Biotechnology, 9, 703241.

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Characterizing Mesenchymal Stem Cells by Flow Cytometry

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Standard flow cytometry was performed to determine the presence of MSC markers, as defined by the International Society for Cellular Therapy.¹⁵ UC‐MSCs were stained using a Human MSC Analysis kit (BD Biosciences, Franklin Lakes, NJ, USA) containing the following mouse monoclonal antibodies, which are positive markers for MSCs: fluorescein isothiocyanate (FITC)‐conjugated anti‐human CD90; phycoerythrin (PE)‐conjugated anti‐human CD105; allophycocyanin (APC)‐conjugated anti‐human CD73; FITC‐conjugated anti‐human CD44 (BD Biosciences), and PE‐conjugated anti‐HLA‐ABC (BD Biosciences). Additionally, UC‐MSCs were stained with FITC‐conjugated anti‐HLA‐DR (BD Biosciences), FITC‐conjugated anti‐human CD34 (BD Biosciences), PE‐conjugated anti‐human CD11b (BD Biosciences), PE‐conjugated anti‐human CD19 (BD Biosciences), or APC‐conjugated anti‐CD45 (BD Biosciences); these are negative markers for MSCs. Propidium iodide was used to identify and exclude dead cells using flow cytometry, as previously described.¹⁶

Tsuji S., Mukai T., Tsuchiya H., Iwatani C., Nakamura A., Nagamura‐Inoue T, & Murakami T. (2023). Impact of administering umbilical cord‐derived mesenchymal stem cells to cynomolgus monkeys with endometriosis. Reproductive Medicine and Biology, 22(1), e12540.

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Phenotypic Characterization of MSCs

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Characterization of hAMSCs and hUC-MSCs phenotype was performed by means of flow cytometry. In passage 4-7, MSCs were seeded in well with Alpha Minimum Essential Medium (αMEM) (Sigma-Aldrich, St. Louis, MO, USA). Afterwards, were fixed with 10% formaldehyde and incubated using the Human MSC Analysis Kit (BD Bioscience, USA) with the addition of a CD90, CD105 and CD73 and negative CD45 cocktail primary antibodies. The primary antibody was labeled using Fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse antibody (Sigma-Aldrich, St. Louis, MO, USA). The cells were then viewed and analyzed by Fluorescence Assisted Cell Sorting (FACS) Calibur flow cytometer (BD Bioscience, USA).

Hendrijantini N, & Hartono P. (2019). Phenotype Characteristics and Osteogenic Differentiation Potential of Human Mesenchymal Stem Cells Derived from Amnion Membrane (HAMSCs) and Umbilical Cord (HUC-MSCs). Acta Informatica Medica, 27(2), 72-77.

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Flow Cytometry Analysis of hUCMSCs and M2 Macrophages

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The cultured hUCMSCs were washed with PBS; then, 1 × 10⁶ hUCMSCs were placed in a centrifuge tube and treated with Human MSC Analysis Kit (562245, BD, USA). After 30 min of maintenance in the dark at 4 °C, hUCMSCs were washed twice by PBS and then resuspended in PBS. These marker proteins were detected by flow cytometry (BD Bioscience, BD FACSCalibur). The same method was used in the measurement of the polarization of M2 macrophages. For detection of specific markers, the suspended macrophages were incubated with antibodies, anti-CD63 (53-5920-80, eBoscience, San Diego, United States) and anti-CD81 (17-2061-82, eBoscience, San Diego, United States) at 4 °C kept in the dark place for 30 min to test the proportion of macrophages.

Li K., Yan G., Huang H., Zheng M., Ma K., Cui X., Lu D., Zheng L., Zhu B., Cheng J, & Zhao J. (2022). Anti-inflammatory and immunomodulatory effects of the extracellular vesicles derived from human umbilical cord mesenchymal stem cells on osteoarthritis via M2 macrophages. Journal of Nanobiotechnology, 20, 38.

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Comparative Analysis of MSC Phenotypes

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Human BMMSCs, ADMSCs, and UCMSCs were cultured according to our previous study [21 (link)]. Briefly, MSCs were seeded in 10 cm culture dishes and cultured in an α-modified Eagle’s medium (α-MEM; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Waltham, MA, USA), penicillin (100 μg/mL), and streptomycin (100 μg/mL) and incubated at 37 °C in a 5% CO₂ atmosphere. The phenotypes of MSCs were analyzed by the flow cytometer, FACS CantoII, using a human MSC analysis kit (562245, BD Company, Franklin Lakes, NJ, USA) [38 (link)].

Chen Y.C., Fu Y.S., Tsai S.W., Wu P.K., Chen C.M., Chen W.M, & Chen C.F. (2022). IL-1b in the Secretomes of MSCs Seeded on Human Decellularized Allogeneic Bone Promotes Angiogenesis. International Journal of Molecular Sciences, 23(23), 15301.

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Phenotypic Characterization of Adipose-Derived Mesenchymal Stem Cells

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To identify human MSCs, the International Society for Cell Therapy (ISCT) has recommended a panel of cell surface markers. AT-MSCs should be positive for CD44, CD73, CD90 and CD105, but negative for CD34, CD45, CD11b or CD14, CD19 and HLA-DR [22 (link)]. Therefore, the phenotype of hAT-MSCs cells was evaluated by flow cytometry using a commercially available kit for human Mesenchymal Stem Cells analysis (Human MSC Analysis Kit, BD Biosciences, USA).
For this purpose, 2.5 × 10⁵ AT-MSCs, from the third passage, were washed twice with cold PBS solution and centrifuged at 4°C for 5 min. Then cells were suspended in Dulbecco’s Phosphate-Buffered Saline (DPBS, Corning, USA) with the addition of 2% Foetal Bovine Serum (FBS) and incubated in the dark for 30 min with antibodies. The cells were then washed twice in PBS, suspended in PBS and analysed with a flow cytometer (BD FACSCanto II, USA).

Siedlecka N., Mecinska-Jundzill K., Fierek E., Fekner Z., Jundzill A., Rasmus M., Kloskowski T., Szeliski K., Bienkowski W., Czajkowski R., Drewa T, & Pokrywczynska M. (2020). Biostimulative effect of laser on growth of mesenchymal stem/stromal cells in vitro. Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii, 37(5), 771-780.

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