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Related Experiment Videos

Decrease of L-selectin expression on human CD34+ cells on freeze-thawing and rapid recovery with short-term

Y Hattori1, H Kato, M Nitta

  • 1Department of Transfusion Medicine, Aichi Medical University, 21 Karimata, Yazako, Aichi 480-1195, Japan.

Experimental Hematology
|February 13, 2001
PubMed
Summary

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Cryopreservation significantly reduces L-selectin expression on hematopoietic progenitor cells (HPCs). However, this loss is reversible with serum incubation, suggesting potential in vivo restoration after transplantation.

Area of Science:

  • Hematology
  • Cell Biology
  • Cryobiology

Background:

  • Hematopoietic progenitor cells (HPCs) home to bone marrow via adhesion molecules.
  • Cryopreservation can alter HPCs, potentially affecting their function.
  • Understanding these alterations is crucial for successful cell transplantation.

Purpose of the Study:

  • To investigate the impact of cryopreservation on adhesion molecule expression in HPCs.
  • To determine the mechanisms behind L-selectin expression changes.
  • To assess the reversibility of these changes and their functional implications.

Main Methods:

  • Flow cytometry and immunofluorescence to analyze adhesion molecule expression on CD34+ cells.
  • Assessment of L-selectin expression after freeze-thawing, DMSO incubation, and serum culture.

Related Experiment Videos

  • Transmigration assays to evaluate the functional relationship between expression and function.

    • Main Results:

    • Freeze-thawing significantly decreased L-selectin expression on HPCs from various sources (MPB, BM, CB).
    • Dimethyl sulfoxide (DMSO) induced L-selectin shedding mediated by matrix metalloproteinase (MMP).
    • Serum incubation, especially with HS-27A stromal cells, restored L-selectin expression to baseline or higher levels.

    Conclusions:

    • Cryopreservation induces reversible loss of L-selectin on HPCs, primarily via MMP-mediated shedding.
    • Restoration of L-selectin expression is possible in vitro, suggesting potential for in vivo recovery.
    • These findings support the feasibility of successful hematopoiesis restoration post-transplantation despite cryopreservation damage.