Which panel of antibodies could be used for the detection of bone marrow stromal and endothelial cells by flow cytometry?

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  Valentina Mele · Universitätsspital Basel

  Hello! I'm always using CD90, CD73 and CD29 for BM-MSCs (FITC, PE and APC respectively), they are positive for all the 3. If you have to discriminate BM-MSCs from other kind of cells you can use CD90, CD45 and CD34, they should be positive for CD90 and negative for the other 2 markers. For endothelial cells I use CD105 and CD31 (FITC and PE). I buy the FACS antibody from BD. I've never used kit!

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  Hello! I'm always using CD90, CD73 and CD29 for BM-MSCs (FITC, PE and APC respectively), they are positive for all the 3. If you have to discriminate BM-MSCs from other kind of cells you can use CD90, CD45 and CD34, they should be positive for CD90 and negative for the other 2 markers. For endothelial cells I use CD105 and CD31 (FITC and PE). I buy the FACS antibody from BD. I've never used kit!

  Mar 30, 2012
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  Efstathios Kastritis · National and Kapodistrian University of Athens

  Thank you so much for your input!

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  Thank you so much for your input!

  Mar 30, 2012
• 
  Andrew Filer · University of Birmingham

  Valentina sounds wise! Just be aware that Endothelial cells will stain positive for CD90 of course, and CD105 is a good marker for MSC as well as EC. So the answer depends upon whether you want to differentiate between MSC and EC, or pick them up as one population. if the former you probably need one more vascular EC marker to be sure. You could use CD31 and CD34 copositivity, or add in von Willebrand factor.

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  Valentina sounds wise! Just be aware that Endothelial cells will stain positive for CD90 of course, and CD105 is a good marker for MSC as well as EC. So the answer depends upon whether you want to differentiate between MSC and EC, or pick them up as one population. if the former you probably need one more vascular EC marker to be sure. You could use CD31 and CD34 copositivity, or add in von Willebrand factor.

  Mar 30, 2012
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  Gopi Kolluru · Louisiana State University Health Sciences Center Shreveport

  Hi! you can use CD133 (APC) CD31 (FITC), Flk1/CD309 (PE) for endothelial cells. We get the antibodies for FACS from BD/ebioscience.

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  Hi! you can use CD133 (APC) CD31 (FITC), Flk1/CD309 (PE) for endothelial cells. We get the antibodies for FACS from BD/ebioscience.

  Mar 30, 2012
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  Christoph Patsch · Rheinische Friedrich-Wilhelms-Universität Bonn

  We use the endothelial specific surface markers CD31 (PECAM-1) and CD144 (VE-Cadherin). Both antibodies are purchased from BD. Could luck with your experiments.

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  We use the endothelial specific surface markers CD31 (PECAM-1) and CD144 (VE-Cadherin). Both antibodies are purchased from BD. Could luck with your experiments.

  Apr 1, 2012
• 
  Pachaiyappan Viswanathan ·

  Don't go by kits, select antibody markers based on your requirment and see for there compatibility in your machine. But using two or three markers you cant clearly define a population. For example, CD34 expressed both on HScs and Endothelial cells, so in this case you have to use markers which are specific to Endothelial lineage and negative to Hematopeoitic lineage. Bone marrow stromal cells are CD 73, CD105, CD90, CD 166 positive and CD 34, CD45, CD14, CD19 And CD133 negative

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  Don't go by kits, select antibody markers based on your requirment and see for there compatibility in your machine. But using two or three markers you cant clearly define a population. For example, CD34 expressed both on HScs and Endothelial cells, so in this case you have to use markers which are specific to Endothelial lineage and negative to Hematopeoitic lineage. Bone marrow stromal cells are CD 73, CD105, CD90, CD 166 positive and CD 34, CD45, CD14, CD19 And CD133 negative

  May 20, 2012
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  Anet Valdés · Center of Molecular Immunology

  For the characterization of CECs you can use CD45, CD31 and CD146 and also the 7AAD marker for distinguish the apoptotic of the viable CECs, but these are realted with the need of your project; for example in my case is for evaluate the effect of an anti-angiogenic droug (in this case you will need 4 cytometer chanels)

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  For the characterization of CECs you can use CD45, CD31 and CD146 and also the 7AAD marker for distinguish the apoptotic of the viable CECs, but these are realted with the need of your project; for example in my case is for evaluate the effect of an anti-angiogenic droug (in this case you will need 4 cytometer chanels)

  May 28, 2012
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  Behzad Kharabi · Children's Hospital Los Angeles

  We had the same discussion during the Cancer Cell revision. You can use MSC markers as mentioned by the others but you should be aware that MSCs are only a small fraction of the bone marrow stroma. Thereby it's difficult to argue that by using MSC markers you reflect the whole BMSCs. Use CD45 first since BMSCs are CD45 negative and later on additional markers.You should also be aware that certain stromal cell populations are not detectable by FACS. But if you need to pin point it down you may consider gene expression profiling. You can check for osteoblastic/osteogenic genes and check later on their expression. Of course you need to sort. CD133 is not a typical endothelial marker. CD31, VE-Cadherin are.

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  We had the same discussion during the Cancer Cell revision. You can use MSC markers as mentioned by the others but you should be aware that MSCs are only a small fraction of the bone marrow stroma. Thereby it's difficult to argue that by using MSC markers you reflect the whole BMSCs. Use CD45 first since BMSCs are CD45 negative and later on additional markers.You should also be aware that certain stromal cell populations are not detectable by FACS. But if you need to pin point it down you may consider gene expression profiling. You can check for osteoblastic/osteogenic genes and check later on their expression. Of course you need to sort. CD133 is not a typical endothelial marker. CD31, VE-Cadherin are.

  Jul 3, 2012