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Stem Cells

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

Leukemia volume 24, pages 153–161 (2010)Cite this article

Abstract

A deeper understanding of stem cell niche engagement and subsequent behaviors would be enhanced by technologies enabling the tracking of individual stem cells at the clonal level in long-term co-culture (LTC), which mimics the complexity of the bone marrow microenvironment in vivo. Here, we report the application of time-lapse imaging with intermittent fluorescence for tracking well-defined populations of GFP+ murine hematopoietic stem cells (HSCs) using LTC for >5 weeks. Long-term (LT) and short-term (ST) repopulating HSCs and hematopoietic progenitor cells (HPCs) were compared. The transition from cobblestone areas (CAs) under the stromal cell mantle into dispersed migrating cells on top of the stroma (COS) were directly observed. The ST-HSC and LT-HSC were able to initiate multiple waves of CA formation and COS expansion beyond 2 and 4 weeks, respectively. Retrospective tracking of individual CA forming cell (CAFC) revealed a preference for residing under stroma before the first division and a longer interval before first division for LT-HSC. Inability to maintain quiescence in subsequent divisions was revealed. Our study represents an important starting point from which the LTC system can be augmented to provide a better in vitro model for bone marrow stem cell niches.

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Acknowledgements

We thank Dr Sallie Boggs for helpful discussion and stimulating suggestions, and Julie Goff for help collecting data. We thank Charalambos Athanassiou and Lei Qian for maintaining the automatic system and for statistical analysis. This investigation was supported by the NIH grants RO1 EB 001051 (to RH), R01 AI080424 and RO1 HL 075601 (to TC). TC was a recipient of the Scholar Award from the Leukemia & Lymphoma Society (1027-09), a recipient of the Changjiang Scholarship from the Ministry of Education of China (2007-JGT-08) and a recipient of the Outstanding Young Scholar Award from the National Natural Science Foundation of China (30825017).

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Author notes

  1. A Bahnson & D Koebler

    Present address: 6Current address: Kairos Instruments, LLC, 520 William Pitt Way, Pittsburgh, PA 15238, USA.,

  2. Y Song and A Bahnson: These two authors contributed equally to this work.

Authors and Affiliations

  1. Department of Radiation Oncology, University of Pittsburgh School of Medicine, Cancer Stem Cell Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA, USA

    Y Song, H Yu, H Shen & T Cheng

  2. Department of Hematology, Huashan Hospital, Fudan University, Shanghai, China

    Y Song & Y Xie

  3. Automated Cell, Inc. 390 William Pitt Way, Pittsburgh, PA, USA

    A Bahnson, N Hall, D Koebler & R Houck

  4. Institute of Hematology, Chinese Academy of Medical Sciences and Peking Union Medical College, State Key Laboratory for Experimental Hematology, Tianjin, China

    T Cheng

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Correspondence to T Cheng.

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Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu)

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Song, Y., Bahnson, A., Hall, N. et al. Stem cell traits in long-term co-culture revealed by time-lapse imaging. Leukemia 24, 153–161 (2010). https://doi.org/10.1038/leu.2009.191

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