Hematopoietic stem cells(HSCs)self-renew or differentiate through division.Cytokines are essential for inducing HSC division,but the optimal cytokine combination to control self-renewal of HSC in vitro remains unclear...Hematopoietic stem cells(HSCs)self-renew or differentiate through division.Cytokines are essential for inducing HSC division,but the optimal cytokine combination to control self-renewal of HSC in vitro remains unclear.In this study,we compared the effects of interleukin-12(IL-12)and thrombopoietin(TPO)in combination with stem cell factor(SCF)on in vitro self-renewal of HSCs.Single-cell assays were used to overcome the heterogeneity issue of HSCs,and serum-free conditions were newly established to permit reproduction of data.In single-cell cultures,CD150^(+)CD48^(-)CD41^(-)CD34^(-)c-Kit^(+)Sca-1^(+)lineage^(-)SCs divided significantly more slowly in the presence of SCF+IL-12 compared with cells in the presence of SCF+TPO.Serial transplantation of cells from bulk and clonal cultures revealed that TPO was more effective than IL-12 at supporting in vitro self-renewal of short-term(<6 months)HSCs,resulting in a monophasic reconstitution wave formation,whereas IL-12 was more effective than TPO at supporting the in vitro selfrenewal of long-term(>6 months)HSCs,resulting in a biphasic reconstitution wave formation.The control of division rate in HSCs appeared to be crucial for preventing the loss of self-renewal potential from their in vitro culture.展开更多
基金supported by grants from the National Key Research and Development Program of China Stem Cell and Translational Research(2017YFA0104903,2016YFA0100600,and 2017YFA0103400)the Ministry of Science and Technology of China(2015CB964403 and,2011CB964801)+1 种基金the CAMS Initiative for Innovative Medicine(2016-I2M-1-017 and 2017-I2M-1-015)the National Natural Science Foundation of China(81470279,81670105,81421002,81400077,and 81500085).
文摘Hematopoietic stem cells(HSCs)self-renew or differentiate through division.Cytokines are essential for inducing HSC division,but the optimal cytokine combination to control self-renewal of HSC in vitro remains unclear.In this study,we compared the effects of interleukin-12(IL-12)and thrombopoietin(TPO)in combination with stem cell factor(SCF)on in vitro self-renewal of HSCs.Single-cell assays were used to overcome the heterogeneity issue of HSCs,and serum-free conditions were newly established to permit reproduction of data.In single-cell cultures,CD150^(+)CD48^(-)CD41^(-)CD34^(-)c-Kit^(+)Sca-1^(+)lineage^(-)SCs divided significantly more slowly in the presence of SCF+IL-12 compared with cells in the presence of SCF+TPO.Serial transplantation of cells from bulk and clonal cultures revealed that TPO was more effective than IL-12 at supporting in vitro self-renewal of short-term(<6 months)HSCs,resulting in a monophasic reconstitution wave formation,whereas IL-12 was more effective than TPO at supporting the in vitro selfrenewal of long-term(>6 months)HSCs,resulting in a biphasic reconstitution wave formation.The control of division rate in HSCs appeared to be crucial for preventing the loss of self-renewal potential from their in vitro culture.
