Session: SUN 303-321-Cancer in Endocrine Tissues
Bench to Bedside
Poster Board SUN-304
Methods:Bone marrow stromal cells were isolated from tail vertebrae before tumor development; cells were cultured with MSC-qualified FBS to maintain their multipotency. FACS was used for identification and isolation of aBSCs from culture based on a four-color selection panel with two positive (CD106 and CD146) and two negative (CD11b and CD45) markers. Colony formation unit (CFU) and differentiation assays were used as evidence for multipotency of the cells after sorting; microarray is used for transcriptomic profiles comparison among these cells from mice with different genotypes.
Results: Cell cultures from tail vertebrae of mice with different genotypes were established. Different percentages of aBSCs were found in mice with different genotypes (non-detectable for WT, 0.777% for Prkar1a+/-, 3.78% for Prkar1a+/-Prkaca+/- and 12.9% for Prkar1a+/-Prkar2a+/-); RNA was extracted from these cells and submitted for transcriptomic analysis. After FACS sorting, cell cultures were established; different colony formation capacities of aBSCs from mice with different genotypes were indicated by CFU, based on the number of colonies formed every 10,000 cells.
Conclusions: Small changes in PKA types and/or activity caused by deficiencies of different PKA subunits appear to activate different populations of bone lesion-initiating cells; the lesions are consequently different both in number and in histology. These studies shed light in how cAMP-dependent PKA controls multipotency of bone marrow stromal cells in adult skeleton and may lead to tumors if aberrant.
Nothing to Disclose: SL, CP, SSS, KMT, ES, MVN, MDLLS, CAS
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