Excessive Extramedullary Hematopoiesis in Cbfa1-Deficient Mice with a Congenital Lack of Bone Marrow

Abstract

Cbfa1, a transcriptional factor that belongs to the runt-domain gene family, plays a pivotal role in osteogenesis. Cbfa1-deficient mice completely lacked both intramembranous and endochondral bone formation owing to the maturational arrest of osteoblasts, resulting in a total lack of bone marrow throughout the entire skeleton. To investigate the development of hematopoiesis in Cbfa1-deficient mice, hematopoietic tissues and peripheral blood were examined. Colony-forming assays of yolk sac at embryonic day 10.5 (E10.5) and liver at E12.5 showed normal numbers of hematopoietic precursors in the mutant embryos. Further, histological examination and flow-cytometric analysis showed normal hematopoietic development of liver and spleen in the mutant embryos until E17.5. However, mutant embryos at E18.5 showed large hematopoietic foci in the periportal area of liver and mild splenomegaly with dilated vessels, and an increase of myeloid cells was observed in both organs. Flow-cytometric analysis also demonstrated a relative increase of granulocytes and a relative decrease of B-cells reciprocally in the livers and spleens of the mutant embryos. Colony-forming assays showed that the frequency of colony-forming unit-culture (CFU-C) was elevated in liver, and both the frequency and total number of CFU-C were increased in spleen of mutant embryos at E18.5. Moreover, leukoerythroblastosis was observed in peripheral blood of the mutant embryos. These data demonstrate that the congenital lack of bone marrow caused excessive levels of extramedullary hematopoiesis in both liver and spleen at the late embryonic stage.