Investigation of multipotent cells in bone lesions of mice with PKA defects

Wednesday, October 10, 2012 — Poster Session II
Noon – 2:00 p.m	Natcher Conference Center, Building 45	NICHD	CANCER-21

Authors

• SS Liu
• C Petrovas
• S Spath
• E Saloustros
• E Makareeva
• M Nesterova
• M De La Luz Sierra
• C.A. Stratakis

Abstract

A mouse model double heterozygous for prkar1a and prkaca (Prkar1a+/- Prkaca+/-) has been generated previously. Osseous lesions resembling fibrous dysplasia are found in the long bone and vertebrae of the mice; cells from these lesions arise from an area proximal to the growth plate, have pre-osteoblast nature, signature of mesenchymal-to-epithelial transition and increased wnt signaling. The hypothesis is a specific population of adult bone stromal cells (aBSCs) are stimulated by alternative PKA activity and give rise to the lesions. We are interested in isolation, cultureation and identification of the aBSCs, to understand the effect of cAMP signaling on mesenchymal cell growth and malignant transformation. Cells either from none lesion culture of 6 month old mice or directly isolation from tail vertebrae bone marrow are investigated. Colony formation assay has also been done, showing presence of cells with colony formation capacity in the culture.FACS is used for identification of aBSCs. Side population assay is used to identify progenitor cells, 4 color FACS experiment with two negative and two positive MSC antibodies is used for identification of MSC from the culture. After sorting, the transcriptomic information of target cells from different genotypes will be compared.

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