Generation of Engraftable Hematopoietic Stem Cells From Induced Pluripotent Stem Cells by Way of Teratoma Formation

Abstract

In vitro generation of hematopoietic stem cells (HSCs) from induced pluripotent stem cells (iPSCs) has the potential to provide novel therapeutic approaches for replacing bone marrow (BM) transplantation without rejection or graft versus host disease. Hitherto, however, it has proved difficult to generate truly functional HSCs transplantable to adult host mice. Here, we demonstrate a unique in vivo differentiation system yielding engraftable HSCs from mouse and human iPSCs in teratoma-bearing animals in combination with a maneuver to facilitate hematopoiesis. In mice, we found that iPSC-derived HSCs migrate from teratomas into the BM and their intravenous injection into irradiated recipients resulted in multilineage and long-term reconstitution of the hematolymphopoietic system in serial transfers. Using this in vivo generation system, we could demonstrate that X-linked severe combined immunodeficiency (X-SCID) mice can be treated by HSCs derived from gene-corrected clonal iPSCs. It should also be noted that neither leukemia nor tumors were observed in recipients after transplantation of iPSC-derived HSCs. Taken our findings together, our system presented in this report should provide a useful tool not only for the study of HSCs, but also for practical application of iPSCs in the treatment of hematologic and immunologic diseases.

Interspecies chimeras for human stem cell research

“…In this system, the contribution of human cells to the peripheral blood was generally very low, and thus it might be worth considering strategies to ʻempty' the host hematopoietic stem cell (HSC) niche prior to transplantation, as others have shown in the field of interspecies organ generation (Kobayashi et al, 2010). Suzuki et al (2013) showed that human HSCs can be produced from human induced pluripotent stem cells (iPSCs) by in vivo teratoma formation in mouse. Since the production of bona fide HSCs from human iPSCs has not yet been achieved in vitro, these data are interesting as they might suggest that using the in vivo biological environment could provide an advantage over in vitro culture.…”

Human haematopoietic stem cell development: from the embryo to the dish

“…The mixture of cell types in these tumours can preserve some developmentally important inductive interactions, leading to the formation of disorganised tissues and organs, including bone marrow (Hentze et al, 2009;Amabile et al, 2013). Teratomas formed from hPSCs engrafted into immunodeficient mice, in some cases, generate human CD45 + cells that home to bone marrow, circulate in the recipient peripheral blood and colonise lymphoid tissues (Amabile et al, 2013;Suzuki et al, 2013). Transplantation of total bone marrow or sorted human CD34 + CD45 + bone marrow cells from primary into secondary recipients shows donor-derived multilineage peripheral blood chimerism, comparable to UCB repopulation (Amabile et al, 2013;Suzuki et al, 2013).…”

“…Teratomas formed from hPSCs engrafted into immunodeficient mice, in some cases, generate human CD45 + cells that home to bone marrow, circulate in the recipient peripheral blood and colonise lymphoid tissues (Amabile et al, 2013;Suzuki et al, 2013). Transplantation of total bone marrow or sorted human CD34 + CD45 + bone marrow cells from primary into secondary recipients shows donor-derived multilineage peripheral blood chimerism, comparable to UCB repopulation (Amabile et al, 2013;Suzuki et al, 2013). Teratomaderived B and T cells appear functionally normal, and erythrocytes express adult haemoglobins, which is not usually achievable during in vitro hPSC differentiation.…”

Long‐term skin regeneration in xenografts from iPSC teratoma‐derived human keratinocytes

“…This is particularly the case for multilineage tissues such as hematopoietic tissue and epidermis (5). The inability to recreate in vitro the microenvironment/niche using specific cytokines and feeder cells led researchers in the hematopoietic field to use teratomas as an alternative source of engraftable functional progenitors (7,8). We thus sought a similar approach for keratinocytes.…”