Prevention of genetic anemias in mice by microinjection of normal hematopoietic stem cells into the fetal placenta

Abstract

Mice homozygous for mutant genes at the W locus have a marked macrocytic anemia that is fatal in some genotypes. The defect is believed to originate in the developmentally pluripotent hematopoietic stem cell population. Anemia is first grossly manifest on day 13 of gestation, when the liver is the chief hematopoietic organ.

Humoral Immune Response to Recombinant Adenovirus and Adeno-Associated Virus after In Utero Administration of Viral Vectors in Mice

“…Induction of tolerance across allogeneic barriers after in utero administration of HSC has been evaluated in a few murine models (22)(23)(24)(25). Significant and durable engraftment of MHC-mismatched HSC has been achieved with HSC transplantation between d 11 and 13 p.c (23).…”

Temporally regulated expression patterns following in utero adenovirus-mediated gene transfer

“…Other methods of fetal gene transfer have included: ex vivo retrovirus-transduced fetal hematopoietic cells which were re-infused in the donor fetus with intermittent detection of transgene activity post-natally; 14 intraperitoneal, retrovirus injection into fetal and adult rats; 15 intraplacental injection at a single developmental time-point; 16 and introduction of normal fetal liver cells or retrovirus-transduced cultured placental cells into the fetal placenta. 17,18 Only a limited number of studies have allowed for development to term and even fewer studies have assessed gene expression longer than a week following vaginal delivery. 9,10,14,15 As the route and timing of administration have been found to be critical determinants of transduction efficiency in neonatal and adult rats, 12,19,20 these variables are likely to be influential for in utero gene transfer as well.…”

Generation of human/rat xenograft animal model for the study of human donor stem cell behaviorsin vivo

“…Human-rat hybrid animals were generated as described by Fleischman and Mintz [11] , with minor modifications. Briefly, micropipettes were prepared from capillary tubing with a vertical pipette puller, and then broken by hand with jewelers' forceps under a dissecting microscope to produce a tip 20-30 gm in diameter with a sharp-pointed bevel.…”