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Introduction on six view points

  1. Top of page
  2. Introduction on six view points
  3. References

Cell-derived microparticles (MPs) are receiving increasing attention in recent years, both as a diagnostic aid and investigative tool [1–4]. Because they carry markers of the parent cell, including those induced by activation or apoptosis, endothelial MPs (EMPs) can provide valuable information on the status of the parent cell, obtainable in no other way. In addition, there is a growing belief that MPs can function as important diffusible vectors of specific adhesins and cytokines promoting cellular interactions and signal transmission [2].

Thus MP analysis constitutes a new avenue for investigation of pathologies in various diseases. Although still considered investigational [1–4], recent results from several laboratories suggest that MP analysis may be poised to enter the mainstream of clinical testing.

However, a major impediment to that end is the wide variety of methodologies used by different laboratories in this field, few of which can be directly compared to the others, and results from which are sometimes inconsistent or conflicting.

As a first step in addressing that problem, the Editor has organized this Forum article, consisting of a brief description of the preferred methods and rationality from each of six active laboratories in the field, including our own [5–10]. Table 1 lists some key features of the six methodological approaches.

Table 1.  Highlights of six methods
 Principle techniqueQuantitationAnti- coagulantPrepare PPPCell-specific identificationPlateletEndothelialLeukocyte
Isolation MP pelletGeneric MP detection
Biro et al.[5]Flow cytometryCountsCitrate1550 × g, 20 min18 000 × g,  30 minAnnexin VCD62P, CD61, CD63CD31, CD62E or CD144CD4, CD8,etc. See Table 1
Dignat-. George et al. [6]Flow cytometryCountsCitrate1500 × g, 15 min 13 000 × g, 2 minAnnexin VCDCD51, CD144 or CD146CD45
Freyssinet et al. [2]Solid phase captureProthromb- inase captureCitrate1500 × g, 15 min 13 000 × g, 2 minAnnexin V, tissue factorCD62P or GPIbaCD31 or  CD62ECD45
Jimenez et al. [10]Flow cytometryCountsCitrate200 × g, 10 min 1500 × g, 7 minCD41 or CD42b & CD31CD31+/ CD42–  or CD62ECD45
Nomura [8]ELISAStandard PMPEDTA1500 × g, 20 minGP IX (capture) CD62P, CD40L
Shet et al.[9]Flow cytometryCountsCitrate13 000 × g,  10 min100 000 × g,  60 minAnnexin VCD41aCD144CD14 (monocyte)

It is seen that major differences exist in the preparation of the MP samples (such as centrifugation), whether or not they are first sedimented and resuspended, means of generic MP detection (4 of 6 use annexin V), and cell lineage-specific antigenic markers. These differences probably account for some of the different findings among the groups.

References

  1. Top of page
  2. Introduction on six view points
  3. References
  • 1
    VanWijk MJ, VanBavel E, Sturk A, Nieuland R. Microparticles in cardiovascular diseases (Review). Cardiovasc Res 2003; 59: 27787.DOI: 10.1016/S0008-6363(03)00367-5
  • 2
    Freyssinet JM. Cellular microparticles: what are they bad or good for? J Thromb Haemost 2003; 1: 165562.DOI: 10.1046/j.1538-7836.2003.00309.x
  • 3
    Horstman LL, Ahn YS. Platelet microparticles: a wide-angle perspective (Review). Crit Rev Oncol/Hematol 1999; 30: 11142.
  • 4
    Horstman LL, Jy W, Jimenez JJ, Ahn YS. Endothelial microparticles as markers of endothelial dysfunction. Front Biosci 2004; 9: 111835.
  • 5
    Biró É, Nieuwland R, Sturk A. Measuring circulating cell-derived microparticles. J Thromb Haemost 2004; 2: 18434.
  • 6
    Dignat-George F, Sabatier F, Camoin-Jau L, Sampol J. Measuring circulating cell-derived microparticles. J Thromb Haemost 2004; 2: 18445.
  • 7
    Hugel B, Zobairi F, Freyssinet J-M. Measuring circulating cell-derived microparticles. J Thromb Haemost 2004; 2: 18467.
  • 8
    Nomura S. Measuring circulating cell-derived microparticles, J Thromb Haemost 2004; 2: 18478.
  • 9
    Shet AS, Key NS, Hebbel RP. Measuring circulating cell-derived microparticles. J Thromb Haemost 2004; 2: 184850.
  • 10
    Jimenez JJ, Jy W, Horstman LL, Ahn YS. Measuring circulating cell-derived microparticles, J Thromb Haemost 2004; 2: 185051.