Organized Development from Human Embryonic Stem Cells after Injection into Immunodeficient Mice

Abstract

Information concerning the development and differentiation of human embryonic stem (hES) cells in vivo is limited. The present study has focused on the in vivo outcome and differentiation of the hESC line HS181, after injection into SCID/beige mice. hES cell-derived teratomas were explored using histological evaluation and by the identification of markers for differentiated cells and tissues. The analyses identified predominant differentiation along a neuronal lineage, the formation of bone/cartilage and epithelia. Fluorescent in situ hybridization (FISH) analysis with a human-specific probe showed the teratomas to be mainly of human origin, with the most organized areas being exclusively human. Importantly, the study revealed interactions between mouse and human tissues, most notably in the formation of vessels. Both mouse and human cells contributed to specific microstructures in which mouse cells could be observed to take on the appropriate histiotypic appearance. Hence, HS181 cells were able to develop into defined mature tissues, supporting the relevant use of this hES cells model for studies of early human development, given the use of appropriate controls for host contribution. Although extensive mitotic activity implicated progenitor cell activity, no detectable multipotent or malignant areas were observed during the observation period. Persisting undifferentiated hESC were not detected.

Neuroblastoma cells injected into experimental mature teratoma reveal a tropism for embryonic loose mesenchyme

“…PSCT were allowed to progress for 45 days before injected with NB tumor cells. The variations of germ layer formation in separate HS181 PSCT formed under the present conditions have been described previously in more detail and were in all cases minor (9,15).…”

“…SCID/Beige (C.B.-17/ GbmsTac-scid-bgDF N7; M&B, Ry, Denmark), male mice, 6-8 weeks old, were used. PSCT were generated by injection of 10 4 -10 5 HS181 under the testicular capsule of mice, as previously described (8,9). PSCT were allowed to progress for 45 days before injected with NB tumor cells.…”

“…Although generically referred to as a tumor, others and we have previously reported evidence that pluripotent stem cell-derived teratoma (PSCT) from PSC Neuroblastoma cells injected into experimental mature teratoma reveal a tropism for embryonic loose mesenchyme S. JAMIL 1* , J. CEDERVALL 2* , I. HULTMAN with normal karyotype can be described as a failed embryonic process including increasingly chaotic embryonic tissues and an emerging organoid development (7,(8)(9)(10)(11). Despite lacking a developmental axis this process can show striking similarities to the events in the human embryo, including and often dominated by components of early neural development (9,10). When compared to human embryos at diagnosed gestation stages, we observed minor kinetic deviations in the appearances of several organoid structures in PSCT developed from the well-studied human embryonic stem cell line HS181 (10).…”

In vitro and in vivo differentiation of human embryonic stem cells into retina‐like organs and comparison with that from mouse pluripotent epiblast stem cells

“…As indicated by various investigators (Gerecht-Nir et al, 2004;Gertow et al, 2004;Yokoo et al, 2008), developing tissues or organs have the ability to induce implanted stem cells to differentiate into the same types of tissues or organs into which the stem cells were transplanted. An impressive case for renal promodial-like ducts induced from mouse ES cells was reported (Yamamoto et al, 2005).…”

Lentiviral-Mediated HoxB4 Expression in Human Embryonic Stem Cells Initiates Early Hematopoiesis in a Dose-Dependent Manner but Does Not Promote Myeloid Differentiation

“…H9 cells, passage 82/83, were harvested mechanically prior to implantation, and 1 ϫ 10 5 cells were inoculated beneath the testicular capsule, as described [28]. The animals were sacrificed after 8 weeks, the teratomas were fixed in 4% neutral buffered formaldehyde overnight prior to dehydration through a graded series of alcohols to xylene.…”