Multiorgan engraftment and differentiation of human cord blood CD34 ⁺ Lin ⁻ cells in goats assessed by gene expression profiling

Abstract

To investigate multitissue engraftment of human primitive hematopoietic cells and their differentiation in goats, human CD34 ؉ Lin ؊ cord blood cells transduced with a GFP vector were transplanted into fetal goats at 45-55 days of gestation. GFP ؉ cells were detected in hematopoietic and nonhematopoietic organs including blood, bone marrow, spleen, liver, kidney, muscle, lung, and heart of the recipient goats (1.2-36% of all cells examined). We identified human ␤2 microglobulin-positive cells in multiple tissues. GFP ؉ cells sorted from the perfused liver of a transplant goat showed human insulin-like growth factor 1 gene sequences, indicating that the engrafted GFP ؉ cells were of human origin. A substantial fraction of cells engrafted in goat livers expressed the human hepatocyte-specific antigen, proliferating cell nuclear antigen, albumin, hepatocyte nuclear factor, and GFP. DNA content analysis showed no evidence for cellular fusion. Long-term engraftment of GFP ؉ cells could be detected in the blood of goats for up to 2 yr. Microarray analysis indicated that human genes from a variety of functional categories were expressed in chimeric livers and blood. The human͞goat xenotransplant model provides a unique system to study the kinetics of hematopoietic stem cell engraftment, gene expression, and possible stem cell plasticity under noninjured conditions. hematopoietic stem cell ͉ transplantation ͉ plasticity ͉ microarray

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

“…By making use of this window of opportunity and performing stem cell transplantation at the appropriate pre-immune stage of fetal life, significant levels of multiorgan, multi-tissue, multi-lineage engraftment are very possible, as confirmed by other investigators [7,8] . More importantly, in contrast to human/mouse xenograft model from the immunodeficient mice for the study of stem cell regulation [9] , such human-rat chimeras possess the normal physiological conditions and potential human immune system reconstituted by donor-derived human differentiated cells [13] .…”

“…Currently, by taking advantage of the window of o p p o r t u n i t y t o p e r f o r m h u m a n s t e m c e l l ( h S C ) t r a n s p l a n t a t i o n d u r i n g t h e p r e i m mu n e s t a g e o f development, fetal sheep [5,7] and fetal goat [8] are developed to be a unique and clinically relevant xenograft animal models for assessing the differentiative potential of hSCs in vivo, but from a scientific perspective, large animals are not a suitable model for mechanistic research.…”

“…To address these issues, there are active sustaining efforts to pursue developing human/animal xenograft models (including mice, sheep, goats, monkeys and pigs) to in vivo study the related fundmental mechanisms [2][3][4][5][6][7][8] .…”

Stem cells in liver regeneration, fibrosis and cancer: the good, the bad and the ugly

“…Nevertheless, bone marrow cell therapies in animal models and patients have produced some interesting results. For example, while bone marrow fails to contribute to liver development in neonatal mice [102], impressive numbers of hepatocytes appear to be human cord blood (CD34 − lin − ) -derived when the cord blood is transplanted in to either pre-immune fetal sheep [103] or goats [104]. In animal models, the most promising cells for therapy appear to be mesenchymal stem cells (MSCs).…”

Bone marrow‐derived very small embryonic‐like stem cells: Their developmental origin and biological significance

“…It is well known that CB-derived cells contribute to skeletal muscle (Gang et al, 2004;Brzoska et al, 2006), liver (Kakinuma et al, 2003;Newsome et al, 2003;Di Campli et al, 2004), neural tissue (Buzanska et al, 2002), and myocardium regeneration (Henning et al, 2004;Ma et al, 2005), and more importantly, recent multiorgan engraftment and differentiation has been achieved in goats after transplantation of human CB CD34 ϩ lin Ϫ cells (Zeng et al, 2006a). Furthermore, several groups of investigators described the presence of Oct-4 ϩ , Nanog ϩ , and SSEA-3/4 ϩ stem cells in human CB and umbilical cord matrix (Carlin et al, 2006).…”