Isolation and Quantitation of Lymphocytes

Splenocytes were harvested from allo-and syn-HSCT recipients by gently crushing tissue between frosted glass slides (Fisher Scientific, Pittsburgh, PA, USA) [25 (link)]. BM cells were harvested from the tibia and femur of one hind leg as described before [27 (link)]. Single cell suspensions were prepared by passing the harvested cell suspension through a cell strainer (Becton Dickinson, Franklin Lakes, NJ, USA). Live lymphocytes were quantitated by fluorescence microscopy as previously described [24 (link)].

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Hossain M.S., Kunter G.M., El-Najjar V.F., Jaye D.L., Al-Kadhimi Z., Taofeek O.K., Li J.M, & Waller E.K. (2017). PD-1 and CTLA-4 up regulation on donor T cells is insufficient to prevent GvHD in allo-HSCT recipients. PLoS ONE, 12(9), e0184254.

Publication 2017

frosted glass slides cell strainer

Femur Fluorescence microscopy Lymphocytes Tibia Tissue

Corresponding Organization : Emory University

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Variable analysis

independent variables

Allogeneic hematopoietic stem cell transplant (allo-HSCT) recipients
Syngeneic hematopoietic stem cell transplant (syn-HSCT) recipients

dependent variables

Splenocytes harvested from allo-HSCT and syn-HSCT recipients
Bone marrow (BM) cells harvested from the tibia and femur
Live lymphocytes quantitated by fluorescence microscopy

control variables

Tissue harvested by gently crushing between frosted glass slides
Single cell suspensions prepared by passing the harvested cell suspension through a cell strainer

controls

No positive or negative controls are explicitly mentioned in the provided information.

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