Stem cell activity of porcine c-kit(+) hematopoietic cells.
Le Guern AC, Giovino MA, Abe M, Theodore PR, Qi J, Down JD, Sachs DH, Sykes M, Yang YG.
BioTransplant, Incorporated, Charlestown, Mass., USA; Immerge Bio Therapeutics, Incorporated, Charlestown, Mass., USA; Transplantation Biology Research Center, Surgical Service, Massachusetts General Hospital, Harvard Medical School, Boston, Mass., USA
Objective. A marker for hematopoietic stem cells (HSCs) of pigs, which are considered to be the most suitable donors for clinical xenotransplantation, has not yet been identified. In this study, we examined the HSC activity of porcine c-kit(+) bone marrow cells (BMCs).Methods. The HSC activity of porcine c-kit(+) BMCs was evaluated both in vitro using colony-forming unit (CFU) and cobblestone area-forming cell (CAFC) assays and in vivo in nonobese diabetic/severe combined immunodeficiency transgenic (NOD/SCID-Tg) mice carrying porcine cytokine transgenes.Results. Purified c-kit(+) BMCs were substantially enriched for both CFUs and CAFCs in vitro and their transplantation led to long-term porcine hematopoiesis in vivo in mice. Although porcine chimerism was detectable in the peripheral blood of NOD/SCID-Tg mice receiving porcine c-kit(-) BMCs at early time points after transplantation, the levels were markedly lower than those in mice receiving purified c-kit(+) BMCs (0.2%+/-0.14% vs 7.7%+/-1.6% and 0.17%+/-0.17% vs 5.6%+/-2.1% at weeks 3 and 6, respectively). Importantly, all mouse recipients of porcine c-kit(+) BMCs showed durable multilineage chimerism (>19 weeks), whereas no recipients of porcine c-kit(-) BMCs sustained long-term engraftment. Moreover, porcine HSCs that had engrafted for 19 weeks in the recipients of porcine c-kit(+) BMCs gave rise to clonogenic progenitors in vitro and reconstituted porcine hematopoiesis in secondary recipients.Conclusion. The present study demonstrates that c-kit is an essential marker of both long-term-repopulating HSCs and progenitor cells with early engraftment capacity. |
