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Delta-Like-1 Changes the Immunomodulatory Property of OP9 Cells.

Zhang L, Dang RJ, Yang YM, Cui DC, Li P, Ni YL, Hao T, Wang C, Jiang XX, Fang NZ - Stem Cells Int (2015)

Bottom Line: With stimulation, both OP9 and OP9DL1 showed increased nitrate oxide (NO) production.The NO levels of OP9 were higher than that of OP9DL1 when stimulated with TNFα/IFNγ or LPS/IL4.DL1 overexpression alone changed the properties of OP9 cells in addition to their role in early B cell development.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Animal Genetic Breeding and Reproduction, Yanbian University, Yanji, Jilin 133002, China ; Department of Advanced Interdisciplinary Studies, Institute of Basic Medical Sciences, 27 Taiping Road, Haidian District, Beijing 100850, China ; Department of Biology and Chemical Engineering, Tongren University, Tongren, Guizhou 554300, China.

ABSTRACT
As stromal cells and recently confirmed mesenchymal stem cells, OP9 cells support hematopoiesis stem cell (HSC) differentiation into the B lymphocyte lineage, yet Delta-like-1 (DL1) overexpressing OP9 (OP9DL1) cells promote the development of early T lymphocytes from HSC. However, the immunomodulatory capacity of OP9 or OP9DL1 on mature B and T cell proliferation has not been elucidated. Here, we show that OP9 and OP9DL1 have similar proliferation capacities and immunophenotypes except DL1 expression. Compared with OP9, OP9DL1 displayed more osteogenesis and less adipogenesis when cultured in the respective induction media. Both OP9 and OP9DL1 inhibited mature B and T cell proliferation. Furthermore, OP9 showed stronger inhibition on B cell proliferation and OP9DL1 exhibited stronger inhibition on T cell proliferation. With stimulation, both OP9 and OP9DL1 showed increased nitrate oxide (NO) production. The NO levels of OP9 were higher than that of OP9DL1 when stimulated with TNFα/IFNγ or LPS/IL4. Taken together, our study reveals a previously unrecognized role of OP9 and OP9DL1 in mature B and T cell proliferation. DL1 overexpression alone changed the properties of OP9 cells in addition to their role in early B cell development.

No MeSH data available.


Related in: MedlinePlus

Effect of OP9 or OP9DL1 on differentiation of bone marrow CD34+ cells. OP9 or OP9DL1 cells (5 × 104/well) were plated in 12-well plates with α-MEM medium plus 20% FBS 12 h prior to the addition of BM CD34+ (5 × 105/well). The coculture was started with α-MEM medium plus 20% FBS containing a final concentration of 5 ng/mL each of IL-7 and Flt-3 ligand (Flt-3L). 12 days later, cells were collected for FACS analysis ((a) and (b)), and EBF1 or GATA3 gene expression was determined by real-time PCR after 5-day coculture, respectively ((c) and (d)). (Error bars present the SD of the mean values, ∗∗P < 0.01.)
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fig3: Effect of OP9 or OP9DL1 on differentiation of bone marrow CD34+ cells. OP9 or OP9DL1 cells (5 × 104/well) were plated in 12-well plates with α-MEM medium plus 20% FBS 12 h prior to the addition of BM CD34+ (5 × 105/well). The coculture was started with α-MEM medium plus 20% FBS containing a final concentration of 5 ng/mL each of IL-7 and Flt-3 ligand (Flt-3L). 12 days later, cells were collected for FACS analysis ((a) and (b)), and EBF1 or GATA3 gene expression was determined by real-time PCR after 5-day coculture, respectively ((c) and (d)). (Error bars present the SD of the mean values, ∗∗P < 0.01.)

Mentions: To examine the ability of OP9 and OP9DL1 to support BM CD34+ cell differentiation to B or T lymphoid lineages, coculture experiments of CD34+ cells with OP9 or OP9DL1 were performed. CD34+ cells were isolated from the femurs and tibiae of 2- or 3-week-old mice and were cultured with either OP9 or OP9DL1. The cells were collected and FACS analysis was performed at day 12 of coculture. As shown in Figures 3(a) and 3(b), there were B220+ cells in CD34+ cells with OP9 coculture and CD3+ cells in CD34+ cells with OP9DL1 coculture. Real-time PCR was used to determine the expression of EBF1 (B cell factor) or GATA3 (T cell factor). As expected, high EBF1 expression was found in the OP9 coculture group whereas high GATA3 expression was found in the OP9DL1 coculture group (Figures 3(c) and 3(d)). Similar to previous reports [5, 9], our results suggest that OP9 supports early B lymphocyte lineage development from CD34+ while OP9DL1 promotes early T cell growth.


Delta-Like-1 Changes the Immunomodulatory Property of OP9 Cells.

Zhang L, Dang RJ, Yang YM, Cui DC, Li P, Ni YL, Hao T, Wang C, Jiang XX, Fang NZ - Stem Cells Int (2015)

Effect of OP9 or OP9DL1 on differentiation of bone marrow CD34+ cells. OP9 or OP9DL1 cells (5 × 104/well) were plated in 12-well plates with α-MEM medium plus 20% FBS 12 h prior to the addition of BM CD34+ (5 × 105/well). The coculture was started with α-MEM medium plus 20% FBS containing a final concentration of 5 ng/mL each of IL-7 and Flt-3 ligand (Flt-3L). 12 days later, cells were collected for FACS analysis ((a) and (b)), and EBF1 or GATA3 gene expression was determined by real-time PCR after 5-day coculture, respectively ((c) and (d)). (Error bars present the SD of the mean values, ∗∗P < 0.01.)
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: Effect of OP9 or OP9DL1 on differentiation of bone marrow CD34+ cells. OP9 or OP9DL1 cells (5 × 104/well) were plated in 12-well plates with α-MEM medium plus 20% FBS 12 h prior to the addition of BM CD34+ (5 × 105/well). The coculture was started with α-MEM medium plus 20% FBS containing a final concentration of 5 ng/mL each of IL-7 and Flt-3 ligand (Flt-3L). 12 days later, cells were collected for FACS analysis ((a) and (b)), and EBF1 or GATA3 gene expression was determined by real-time PCR after 5-day coculture, respectively ((c) and (d)). (Error bars present the SD of the mean values, ∗∗P < 0.01.)
Mentions: To examine the ability of OP9 and OP9DL1 to support BM CD34+ cell differentiation to B or T lymphoid lineages, coculture experiments of CD34+ cells with OP9 or OP9DL1 were performed. CD34+ cells were isolated from the femurs and tibiae of 2- or 3-week-old mice and were cultured with either OP9 or OP9DL1. The cells were collected and FACS analysis was performed at day 12 of coculture. As shown in Figures 3(a) and 3(b), there were B220+ cells in CD34+ cells with OP9 coculture and CD3+ cells in CD34+ cells with OP9DL1 coculture. Real-time PCR was used to determine the expression of EBF1 (B cell factor) or GATA3 (T cell factor). As expected, high EBF1 expression was found in the OP9 coculture group whereas high GATA3 expression was found in the OP9DL1 coculture group (Figures 3(c) and 3(d)). Similar to previous reports [5, 9], our results suggest that OP9 supports early B lymphocyte lineage development from CD34+ while OP9DL1 promotes early T cell growth.

Bottom Line: With stimulation, both OP9 and OP9DL1 showed increased nitrate oxide (NO) production.The NO levels of OP9 were higher than that of OP9DL1 when stimulated with TNFα/IFNγ or LPS/IL4.DL1 overexpression alone changed the properties of OP9 cells in addition to their role in early B cell development.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Animal Genetic Breeding and Reproduction, Yanbian University, Yanji, Jilin 133002, China ; Department of Advanced Interdisciplinary Studies, Institute of Basic Medical Sciences, 27 Taiping Road, Haidian District, Beijing 100850, China ; Department of Biology and Chemical Engineering, Tongren University, Tongren, Guizhou 554300, China.

ABSTRACT
As stromal cells and recently confirmed mesenchymal stem cells, OP9 cells support hematopoiesis stem cell (HSC) differentiation into the B lymphocyte lineage, yet Delta-like-1 (DL1) overexpressing OP9 (OP9DL1) cells promote the development of early T lymphocytes from HSC. However, the immunomodulatory capacity of OP9 or OP9DL1 on mature B and T cell proliferation has not been elucidated. Here, we show that OP9 and OP9DL1 have similar proliferation capacities and immunophenotypes except DL1 expression. Compared with OP9, OP9DL1 displayed more osteogenesis and less adipogenesis when cultured in the respective induction media. Both OP9 and OP9DL1 inhibited mature B and T cell proliferation. Furthermore, OP9 showed stronger inhibition on B cell proliferation and OP9DL1 exhibited stronger inhibition on T cell proliferation. With stimulation, both OP9 and OP9DL1 showed increased nitrate oxide (NO) production. The NO levels of OP9 were higher than that of OP9DL1 when stimulated with TNFα/IFNγ or LPS/IL4. Taken together, our study reveals a previously unrecognized role of OP9 and OP9DL1 in mature B and T cell proliferation. DL1 overexpression alone changed the properties of OP9 cells in addition to their role in early B cell development.

No MeSH data available.


Related in: MedlinePlus