The ability for hematopoietic stem cells (HSCs) to regenerate the vascular and blood systems following injury suggests great potential for future therapies. Unfortunately, the various signaling pathways that regulate the regeneration of the adult HSC population in the bone marrow are not clearly understood. One of the proposed regulators for the regeneration of the hematopoietic system is an extracellular secreted protein R Spondin-2 (Rspo2), also known as roof plate-specific spondin-2. The novel interaction between the Rspo2 protein and c-kit+sca-1+lineage– (KSL) HSCs shows an increased number of KSL and of more differentiated hematopoietic stem and progenitor cells (HSPCs) in vivo during both the steady and injured states. To determine the most efficient concentration of Rspo2 for such an interaction to occur, various doses of Rspo2 recombinant protein are plated with the KSL cells. They are then examined through flow cytometry and colony forming cell (CFC) assay. Rspo2 is widely known to interact with the canonical Wnt3a protein to activate the beta-catenin pathway. However, when various concentrations of Rspo2 recombinant proteins are plated with the Wnt3a protein, the results show the opposite effect of plating the cells only with the Rspo2 protein. The overall increase in the total number of cells and KSL cells was concluded to be not significant. This study nonetheless provides the scientific community with a greater foundation for the usage of Rspo2 concentration for future experiments.
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/45572 |
| Date | 03 February 2023 |
| Creators | Jang, Seok Hee Jenny |
| Contributors | Murphy, George J., Chute, John P. |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
| Rights | Attribution 4.0 International, http://creativecommons.org/licenses/by/4.0/ |
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