Human ex vivo hematopoietic stem cell expansion using bone derived reinvigorating mesenchymal stromal cells
Abstract
Many patients lose healthy cells during anti-cancer treatments due to radiation. If successful hematopoietic stem cell transplantation occurs, it would replenish their entire hematopoietic system. HSCs can be utilized in these clinical settings, laboratory work, or gene editing. My research involved culturing HSCs alongside mesenchymal stromal cells, another cell found in the HSC’s origin. Because MSCs are known to regulate and maintain HSCs inside of the bone marrow, I predicted that they may perform the same ex vivo. My students and I plated a monoculture of HSCs and a coculture of MSCs with HSCs to compare, and as hypothesized, the cocultures retained the HSCs, but the cells in the monoculture bursted and died. This research is currently ongoing. Furthermore, passaging cells induce stress, and the population number and characteristics may alter under these conditions. In the cell culture, they are passaged constantly after growing and “age.” I decided to take on my project of Characterizing MSCs, questioning if the cells can still be called “MSCs” after isolation and passages from their characteristic markers. Taking four samples of different passages and patients, I counted, plated, and tracked proliferation, used CFUs to check how many have clonogenic capacities, and checked for various MSC markers with a cytometer. I assumed that one patient’s sample was mishandled previously before freezing. In conclusion, MSCs do not display identical markers after isolation, and passages under ten do not alter their effectiveness as much as cell management.
Human ex vivo hematopoietic stem cell expansion using bone derived reinvigorating mesenchymal stromal cells
TBD
Many patients lose healthy cells during anti-cancer treatments due to radiation. If successful hematopoietic stem cell transplantation occurs, it would replenish their entire hematopoietic system. HSCs can be utilized in these clinical settings, laboratory work, or gene editing. My research involved culturing HSCs alongside mesenchymal stromal cells, another cell found in the HSC’s origin. Because MSCs are known to regulate and maintain HSCs inside of the bone marrow, I predicted that they may perform the same ex vivo. My students and I plated a monoculture of HSCs and a coculture of MSCs with HSCs to compare, and as hypothesized, the cocultures retained the HSCs, but the cells in the monoculture bursted and died. This research is currently ongoing. Furthermore, passaging cells induce stress, and the population number and characteristics may alter under these conditions. In the cell culture, they are passaged constantly after growing and “age.” I decided to take on my project of Characterizing MSCs, questioning if the cells can still be called “MSCs” after isolation and passages from their characteristic markers. Taking four samples of different passages and patients, I counted, plated, and tracked proliferation, used CFUs to check how many have clonogenic capacities, and checked for various MSC markers with a cytometer. I assumed that one patient’s sample was mishandled previously before freezing. In conclusion, MSCs do not display identical markers after isolation, and passages under ten do not alter their effectiveness as much as cell management.
