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The hematopoietic stem cells or HSCs are multipotent, meaning they can differentiate and give rise to all blood and immune cells. HSCs are maintained in the quiescent stage until an external stimulus initiates their differentiation. The multipotent HSCs exist as two heterogeneous populations, long-term repopulating cells (LTRC) and short-term repopulating cells (STRC). The two HSC populations have different surface markers or receptors and are classified based on quiescence and long-term...
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All blood and immune cells are produced from the multipotent hematopoietic stem cells (HSCs) by the process of hematopoiesis. However, they all have a limited life span. In addition, many are depleted in immune surveillance or combatting an injury or infection. This makes blood one of the most regenerative tissues. Hematopoiesis helps replenish these blood and immune cells, restoring the body's normal functioning. However, overproduction of blood and immune cells can make them cancerous or...
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Analysis of hematopoietic stem cells using a composite approach.

Rohit P Nagare1, S Sneha1, S Ramesh2

  • 1Laboratory for Cancer Biology, Cancer Institute (W.I.A), 38, Sardar Patel Road, Chennai, 600 036, Tamilnadu, India.

The International Journal of Biochemistry & Cell Biology
|February 19, 2019
PubMed
Summary

Hematopoietic stem cells (HSCs) were identified using surface markers and functional assays. Results show functional assays identify different cell populations, with both CD34+ and ALDH1A1+ cells in G1 phase, not G0.

Keywords:
ApheresisCell cycle analysisCombined approachFunctional assayHematopoietic stem cellsSurface markers

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Area of Science:

  • Hematology
  • Stem Cell Biology
  • Cancer Research

Background:

  • Cancer stem cells (CSCs) and stem cells are identified using surface markers and functional assays.
  • Functional assays like ALDEFLUOR and side population analysis are marker-independent methods.
  • Uncertainty exists whether these diverse methods identify the same cell populations.

Purpose of the Study:

  • To investigate if surface marker (CD34) and functional assays (ALDEFLUOR, side population) identify consistent cell populations in hematopoietic stem cells.
  • To analyze the cell cycle profile of stem cell populations identified by different methods.

Main Methods:

  • Hematopoietic stem cells (HSCs) and peripheral blood stem cells (PBSCs) were isolated from bone marrow and G-CSF stimulated peripheral blood.
  • Cells were analyzed for CD34 expression combined with ALDEFLUOR and side population assays.
  • Cell cycle analysis was performed using simultaneous labeling with Hoechst and Pyronin Y.

Main Results:

  • CD34+ and CD34- cells co-existed with ALDH1A1+ cells, but side population did not segregate with CD34+ cells.
  • Functional assays identified distinct stem cell populations.
  • Both ALDH1A1+ and CD34+ cells were found in the G1 phase, not the quiescent G0 phase.

Conclusions:

  • Surface marker and functional assays identify different hematopoietic stem cell populations.
  • ALDH1A1 and CD34 expression are associated with cells in the G1 phase of the cell cycle.