Stem cell traits in long-term co-culture revealed by time-lapse imaging

Song, Yifang; Bahnson, Alfred; Hall, Nathan C.; Yu, Hui; Shen, Hongmei; Koebler, Doug; Houck, Raymond K.; Xie, Yi; Cheng, Tao

doi:10.1038/leu.2009.191

Cited by 16 publications

(10 citation statements)

References 37 publications

(38 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the long-term culture of p18 +/+ mouse BMNCs 22, 23 (Fig. 3e), there was no significant difference in CA (cobblestone area) yields in the first 4 weeks between the control group and the groups treated with p18 small-molecular inhibitors.…”

Section: Resultsmentioning

confidence: 94%

Small-molecule inhibitors targeting INK4 protein p18INK4C enhance ex vivo expansion of haematopoietic stem cells

et al. 2015

View full text Add to dashboard Cite

Among cyclin-dependent kinase inhibitors that control the G1 phase in cell cycle, only p18 and p27 can negatively regulate haematopoietic stem cell (HSC) self-renewal. In this manuscript, we demonstrate that p18 protein is a more potent inhibitor of HSC self-renewal than p27 in mouse models and its deficiency promoted HSC expansion in long-term culture. Single-cell analysis indicated that deleting p18 gene favoured self-renewing division of HSC in vitro. Based on the structure of p18 protein and in-silico screening, we further identified novel small-molecule inhibitors that can specifically block the activity of p18 protein. Our selected lead compounds were able to expand functional HSCs in a short-term culture. Thus, these putative small-molecule inhibitors for p18 protein are valuable for further dissecting the signalling pathways of stem cell self-renewal and may help develop more effective chemical agents for therapeutic expansion of HSC.

show abstract

Section: Resultsmentioning

confidence: 94%

Small-molecule inhibitors targeting INK4 protein p18INK4C enhance ex vivo expansion of haematopoietic stem cells

et al. 2015

View full text Add to dashboard Cite

show abstract

“…Ews Ϫ/Ϫ (KO) and Ews ϩ/Ϫ heterozygous (HT)-derived BM stromal cells were equally sufficient as WT cells in providing necessary signals to support CAFC activity ( Figure 7C), suggesting that there were no apparent defects in Ews Ϫ/Ϫ stromal cells in terms of cell adhesion. However, because the CAFC assay does not reflect all aspects of in vivo microenvironment, 38 the impaired hematopoietic activity in Ews Ϫ/Ϫ mice may not necessarily be due solely to an intrinsic alteration of stem cells.…”

Section: Resultsmentioning

confidence: 99%

Ewing sarcoma gene Ews regulates hematopoietic stem cell senescence

Cho

Shen²,

et al. 2011

Blood

View full text Add to dashboard Cite

The longevity of organisms is maintained by stem cells. If an organism loses the ability to maintain a balance between quiescence and differentiation in the stem/ progenitor cell compartment due to aging and/or stress, this may result in death or age-associated diseases, including cancer. Ewing sarcoma is the most lethal bone tumor in young patients and arises from primitive stem cells. Here, we demon- IntroductionEwing sarcoma is an aggressive disease that predominantly afflicts children and young adults, in which cancer cells proliferate in bones and soft tissues. 1 The Ewing sarcoma gene (EWS) encodes an RNA binding protein whose specific functional targets are largely unknown, 2 but abnormal fusion between the EWS N-terminus and the C-terminus of various transcription factors is thought to be the primary cause of Ewing sarcoma. 3 The t(11;22)(q24;q12) translocation, which fuses the EWS and Fli-1 (Friend leukemia virus integration 1) genes, accounts for most cases of Ewing sarcoma. The chromosomal translocation creates a chimeric oncogene encoding a transactivating domain from the EWS protein and the DNA binding domain of Fli-1. 3 EWS-Fli-1 expression results in a bypass of cellular senescence, 4 implying a molecular link between EWS and cellular senescence.Unexpectedly, ectopic expression of EWS-Fli-1 in primary cells or cell lines induces growth arrest or cell death rather than promoting cellular transformation, suggesting that cellular context is critical for the oncogenic potential of EWS-Fli-1. 5 Recent studies have demonstrated that human and mouse bone marrow (BM) mesenchymal stem cells expressing EWS-Fli-1, when engrafted into NOD/SCID mice, induce a malignancy with similar pathologic features as Ewing sarcoma. 6,7 These data indicate that mesenchymal stem cells provide a permissive environment for EWS-Fli-1-mediated transformation, suggesting that Ewing sarcoma occurs at the level of stem/progenitor cells. Consistent with this observation, ectopic expression of an EWS-ERG fusion protein in hematopoietic progenitor cell results in leukemia. 8 In addition, Cre-mediated activation of EWS-Fli-1 in Cre-inducible EWS-Fli-1 mice results in rapid development of myeloid/erythroid leukemia, thereby strengthening the molecular link between this oncogene and hematopoietic malignancy. 3

show abstract

“…As previously described [26], single HSCs of lineage − Sca-1 + c-kit + CD34 − were deposited at one cell per well in 96-well plates with medium containing 100 ng/ml of mouse stem cell factor (mSCF), 10 ng/ml of mouse interleukin 3 (mIL-3), 25 ng/ml of mouse thrombopoietin (mTPO) and 10 ng/ml of mouse erythropoietin (mEPO). Cells were cultured at 37 °C, 5% CO 2 incubator for 10 days.…”

Section: Methodsmentioning

confidence: 99%

Bid is a positive regulator for donor-derived lymphoid cell regeneration in γ-irradiated recipients

Shen¹,

Yu²,

Liang³

et al. 2011

Experimental Hematology

View full text Add to dashboard Cite

Objective Hematopoietic regeneration is regulated by cell survival proteins such as the Bcl-2 family. Bid, a BH3-only protein of the Bcl-2 family, has multiple cellular functions and is involved in a variety of physiological or pathological conditions. We attempted to define its role in hematopoietic cell repopulation under the stress condition of bone marrow transplantation (BMT). Materials and Methods We performed conventional or competitive BMT with donor hematopoietic cells from Bid−/− or Bid+/+ mice. Flow Cytometry was used for quantification of hematopoietic stem cells (HSCs), hematopoietic progenitor cells (HPCs) and differentiated cells in different lineages (T, B and Myeloid cells). Single cell culture and homing assays were performed to further evaluate HSC functions. HPCs were also measured by the colony-forming cell (CFC) culture. Results Contrary to the widely-recognized role of Bid as a pro-apoptotic protein, the absence of Bid significantly reduced the reconstitution of donor hematopoietic cells in γ-irradiated recipients. Interestingly, however, numbers of HSCs and HPCs, and their functions were not overtly altered. Instead, the regeneration of donor T and B cells was significantly impaired in the absence of Bid. Further analysis indicated an accumulation of the triple negative T cell population in the thymus, and pro-B cells in the bone marrow. Conclusion Our current study demonstrates a positive impact of Bid on hematopoietic regeneration mainly due to its unique effects on donor lymphopoiesis in the transplant recipients.

show abstract

Stem cell traits in long-term co-culture revealed by time-lapse imaging

Cited by 16 publications

References 37 publications

Small-molecule inhibitors targeting INK4 protein p18INK4C enhance ex vivo expansion of haematopoietic stem cells

Small-molecule inhibitors targeting INK4 protein p18INK4C enhance ex vivo expansion of haematopoietic stem cells

Ewing sarcoma gene Ews regulates hematopoietic stem cell senescence

Bid is a positive regulator for donor-derived lymphoid cell regeneration in γ-irradiated recipients

Contact Info

Product

Resources

About