Gene expression profile in single cells of neonatal murine anemic liver

In the mice fetus or neonate, the liver is the major hematopoietic organ before the bone marrow takes over, and anemic-associated hypoxia may alter the hematopoietic niche in the liver. Using scRNA-seq analysis, we analyzed the cell diversity in the neonatal murine anemic liver. Briefly, C57BL/6 mice from each litter were randomly assigned to two study groups: (a) naive control and (b) phlebotomy-induced anemia. Phlebotomy-induced anemia was performed in mouse pups by facial vein phlebotomy to remove 20µL blood/g body weight on P2, P4, P6, P8, and P10 to reduce the hematocrit (Hct) index from an initial 45-60% range, down to 20-24%. Hematocrits and RBC indices were measured at each phlebotomy; 5 µL blood was diluted 1:20 in Cellpak reagent (Sysmex America, Mundelein, IL; catalog #DCL-310A), and analyzed in the Sysmex XT-2000iV veterinary hematology analyzer. Non-phlebotomized control animals were handled similarly to phlebotomized animals with a daily non-phlebotomizing needle stick. Pups were weighed daily to determine the volume of blood to be drawn. The animals were euthanized 24 h (P11) later, and the liver tissue was processed for scRNA sequencing.