129in Utero Transplantation of Human Hematopoietic Stem Cells (Hsc) Into Fetal Goats Offers a New Approach to Studies of HSC Expansion and Development

Abstract

In utero transplantation of xenogeneic stem cells into animals (such as mouse, sheep, goats) is a new approach to the studies of the engraftmen, expansion and development of foreign cells in vivo. In addition, such an approach is able to offer a model to clinical potential for correcting or replacing damaged tissues in a variety of disorders, including many that are inherent as well as acquired. Recently, we generated a human/goat hematopoietic stem cell (HSC) xenogeneic model to investigate the engraftment, expansion and development of human hematopoietic cells at an in vivo level. Human HSCs (lin-cells) were isolated and purified from fresh cord blood, and then injected intraperitoneally into pre-immune fetal goats at 55–65 gestation days (term: 145 days). A total of 50 fetal goats were injected. The growth and development of the recipient goats were monitored by B-type ultrasound scan. Ten goats suffered miscarriage due to GVH response; 39 recipients survived a complete gestation and were born alive. All these live-born goats were healthy and grew very well. They could produce offspring normally. In order to determine whether human cells were present longterm in the circulatory blood of the recipient goats, FACS, real-time PCR, RT-PCR, and Southern-blot hybridization as well as FISH were used to analyze the genotype and phenotype up to 11 months after birth. The results showed that hematopoietic chimerism (on average, >108 human cells per goat) was documented in 90% (35/39) of live-born goats that had been originally transplanted with 105 human HSCs. The regenerated human cells included myeloid, B-lymphoid and erythroid lineages as well as more primitive cells as shown by specific positive staining for human CD14, CD20, glycophorin A (GPA) and CD34, and the detection of human-specific DNA sequences and human GPA and CD34 transcripts in the blood of the transplanted goats. No human T or NK-lineage cells were detected. These studies demonstrate the feasibility of using in utero-transplanted fetal goats as a new approach for evaluating the longterm engraftment potential, expansion and development of human hematopoietic cells in vivo.