Cd34 bone marrow cells

Manufactured by Lonza

CD34+ bone marrow cells are a type of hematopoietic stem cells derived from the bone marrow. They express the CD34 surface antigen, which is a marker for stem and progenitor cells. These cells have the potential to differentiate into various blood cell types, including red blood cells, white blood cells, and platelets.

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Lab products found in correlation

C2 point scanning confocal microscope, by Nikon (2 mentions) Smartdish, by STEMCELL (2 mentions) Methocult, by STEMCELL (1 mentions) Methocult h4034 optimum, by STEMCELL (1 mentions) Nis elements viewer, by Nikon (1 mentions)

2 protocols using cd34 bone marrow cells

Co-culture of CAR T cells with CD34+ cells

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Media, GRP78.Δ, GRP78.1x-CAR, or GRP78.2x-CAR T cells were co-cultured with CD34⁺ bone marrow cells (Lonza, Basel, Switzerland) at an E:T ratio of 1:1 and 5:1 for 4 h and were then plated in the presence of MethoCult (Stemcell Technology, Vancouver, CA) in a 6-well SmartDish® (Stemcell Technology, Vancouver, CA), and incubated for 12-14 days at 37 °C. For the CFU assay with GRP78.2x-CAR T cells, NT T cells were also included as a control effector cell population. Plates were imaged using a Nikon C2 point-scanning confocal Microscope (Nikon, Tokyo, Japan) using a ×4 objective. BFU-E (burst Forming Unit—erythroid) and CFU colonies (colony-forming unit—erythroid: CFU-E, colony-forming unit—granulocyte, erythroid, macrophage, megakaryocyte: CFU-GEMM), and colony-forming unit—granulocyte, macrophage + CFU-GM) were enumerated.

Hebbar N., Epperly R., Vaidya A., Thanekar U., Moore S.E., Umeda M., Ma J., Patil S.L., Langfitt D., Huang S., Cheng C., Klco J.M., Gottschalk S, & Velasquez M.P. (2022). CAR T cells redirected to cell surface GRP78 display robust anti-acute myeloid leukemia activity and do not target hematopoietic progenitor cells. Nature Communications, 13, 587.

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Co-culture of CAR T cells and Bone Marrow

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Media, GRP78-CAR, CD123-CAR, or (G4S)3-CAR T cells were co-cultured with CD34⁺ bone marrow cells (Lonza, Basel, Switzerland) at an E/T ratio of 1:1 for 4 h. Cocultures were then plated in the presence of MethoCult H4034 Optimum (Cat#: 04034, Stemcell Technology, Vancouver, CA) in a 6-well SmartDish (Cat#: 27371, Stemcell Technology, Vancouver, CA), and incubated for 12 days at 37°C. Plates were imaged using a Nikon C2 point-scanning confocal Microscope (Nikon, Tokyo, Japan) using a ×4 objective. LIM images from the Nikon software were converted to TIF files by NIS-Elements viewer (5.21, 64-bit, Nikon, Tokyo, Japan). Then analyses were performed manually with FIJI (ImageJ) software.⁷⁰ (link) BFU-E (burst Forming Unit—erythroid) and CFU colonies (colony-forming unit—erythroid: CFU-E, colony-forming unit—granulocyte, erythroid, macrophage, megakaryocyte: CFU-GEMM), and colony-forming unit—granulocyte, macrophage + CFU-GM) were counted and characterized per MethoCult H4034 Optimum standard procedure.

Zoine J.T., Immadisetty K., Ibanez-Vega J., Moore S.E., Nevitt C., Thanekar U., Tian L., Karouni A., Chockley P.J., Arthur B., Sheppard H., Klco J.M., Langfitt D.M., Krenciute G., Gottschalk S., Babu M.M, & Velasquez M.P. (2024). Peptide-scFv antigen recognition domains effectively confer CAR T cell multiantigen specificity. Cell Reports Medicine, 5(2), 101422.

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