Screening of MSC Morphological Responses to Inflammatory Signals to Identify Optimal Conditions for Enhanced Immunosuppressive Function

Project Title: Screening of MSC Morphological Responses to Inflammatory Signals to Identify Optimal Conditions for Enhanced Immunosuppressive Function

 

Project Mentors:

 

 

Project Description:Mesenchymal stromal cells (MSCs) have shown promise in the context of many immune diseases; however, successful clinical translation has not yet been achieved.  In most studies, MSCs are manufactured using standard culture techniques in an environment that does not mimic that experienced by MSCs in diseased tissue (i.e. inflammation).  Stimulation of MSCs with functionally-relevant stimuli, such as inflammatory cytokines, prior to use in in vitro and in vivo models of immune disease has been shown to enhance the ability of MSCs to suppress activated immune cells such as T cells.  Although the effects of these inflammatory signals, such as Interferon-gamma (IFNg) and Tumor Necrosis Factor alpha (TNFa), have been shown to work in different settings there is a need to optimize these preconditioning strategies for enhancing MSC immunosuppressive capacity.  As MSC morphological response to inflammatory signals can be indicative of immunosuppressive capacity, this project aims to use high content imaging to characterize the morphological response of MSCs to a wide range of preconditioning strategies to effectively screen for the optimal concentrations to be used for enhancing MSC immunosuppression of T cells.

 

REU Student Role and Responsibility:The student will be responsible for culturing human MSCs in a high throughput manner to enable screening of large quantities of conditions using high content imaging.  Extensive cell culture, automated liquid handling, high content imaging, and automated image analysis will be performed as part of this project.  The results of this work will be combined with other efforts in the lab to characterize the functional capacity of MSCs stimulated with different preconditions identified from the morphological screen.

 

Required skills or courses for the REU student:The ideal student will have experience with mammalian cell (adherent) cell culture and in vitro cell assays (cell staining, ELISAs, PCR, etc.) Experience with high content imaging and automated image analysis (i.e. CellProfiler) is highly desired, but not required.  The ability to manage and analyze large datasets is also preferred. Students must be entering either their junior or senior year in the Fall 2019 semester and be enrolled in a biomedical/biochemical engineering related field.

 

Expected Outcome for REU student:The outcome of the REU student will be foundational for many future studies performed in the Marklein Lab as the method for screening morphological responses and identifying optimal conditions is relevant to many different cell types and therapeutic strategies beyond MSCs for immune disease treatment.  The data generated form this project will supplement additional data to help support a future publication, as well as likely provide the basis for a conference poster/talk.