EPO-Independent Cell Lines

The EPOR is a required receptor for erythroblast maturation. Using CRISPR Cas9 genome editing, the aim of this project is to create HUDEP2 and KITCAT cells lines that have mutations resulting in constitutively active EPOR. This will decrease cell culture costs but also serve as a model to study EPOR signaling in erythroblasts

Sub Project: Optimization of K562 and KITCAT cell growth conditions: K562 is an established cell line while KITCAT cells are a version of HUDEP recently created in the lab of Laurie Steiner (out collaborators at the U of R). These cell lines are grown in media conditions that can be expensive and hard to maintain. The aim of this project is to determine if alternative reagents and media conditions are sufficient to grow these cell lines. This will allow the lab to increase research focused on human disease leading to anemia using these cell lines as models.

Human erythroblasts with c-Kit activating mutations have reduced cell culture costs and remain capable of terminal maturation - PubMedA major barrier to the in vitro production of red blood cells for transfusion therapy is the cost of culture components, with cytokines making up greater than half of the culture costs. Cell culture cytokines also represent a major expense for in vitro studies of human erythropoiesis. HUDEP-2 cells …

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