Embryonic stem cells as an alternate marrow donor source: engraftment without graft-versus-host disease.
A single embryonic stem cell (ESC) line can be repetitively cryopreserved, thawed, expanded, and differentiated into various cellular components serving as a potentially renewable and well-characterized stem cell source. Therefore, we determined whether ESCs could be used to reconstitute marrow and blood in major histocompatibility complex (MHC)-mismatched mice. To induce differentiation toward hematopoietic stem cells (HSCs) in vitro, ESCs were cultured in methylcellulose with stem cell factor, interleukin (IL)-3, and IL-6. ESC-derived, cytokine-induced HSCs (c-kit+/CD45+) were isolated by flow cytometry and injected either intra bone marrow or intravenously into lethally irradiated MHC-mismatched recipient mice. From 2 wk to 6 mo after injection, the peripheral blood demonstrated increasing ESC-derived mononuclear cells that included donor-derived T and B lymphocytes, monocytes, and granulocytes without clinical or histologic evidence of graft-versus-host disease (GVHD). Mixed lymphocyte culture assays demonstrated T cell tolerance to both recipient and donor but intact third party proliferative responses and interferon gamma production. ESCs might be used as a renewable alternate marrow donor source that reconstitutes hematopoiesis with intact immune responsiveness without GVHD despite crossing MHC barriers.
Burt
Richard K
RK
Division of Immunotherapy, Department of Medicine, Feinberg School of Medicine, Northwestern University, 320 East Superior Street, Searle, Room 3-489, Chicago, IL 60611, USA. rburt@nwu.edu
Verda
Larissa
L
Kim
Duck-An
DA
Oyama
Yu
Y
Luo
Kehuan
K
Link
Charles
C
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002678:Chimera; D003114:Colony-Forming Units Assay; D016207:Cytokines / Q000494:pharmacology; D005260:Female; D006086:Graft vs Host Disease / Q000517:prevention & control; D006410:Hematopoiesis / Q000187:drug effects; D018380:Hematopoietic Stem Cell Transplantation; D007108:Immune Tolerance; D007121:Immunocompetence; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D033581:Stem Cell Transplantation; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology; D039901:Totipotent Stem Cells
D016207:Cytokines