Measuring circulating cell-derived microparticles
Version of Record online: 28 SEP 2004
Journal of Thrombosis and Haemostasis
Volume 2, Issue 10, pages 1842–1843, October 2004
How to Cite
Jy, W., Horstman, L. L., Jimenez, J. J. and Ahn, Y.S. (2004), Measuring circulating cell-derived microparticles. Journal of Thrombosis and Haemostasis, 2: 1842–1843. doi: 10.1111/j.1538-7836.2004.00936.x
- Issue online: 28 SEP 2004
- Version of Record online: 28 SEP 2004
Introduction on six view points
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 .
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.
|Principle technique||Quantitation||Anti- coagulant||Prepare PPP||Cell-specific identification||Platelet||Endothelial||Leukocyte|
|Isolation MP pellet||Generic MP detection|
|Biro et al.||Flow cytometry||Counts||Citrate||1550 × g, 20 min||18 000 × g, 30 min||Annexin V||CD62P, CD61, CD63||CD31, CD62E or CD144||CD4, CD8,etc. See Table 1|
|Dignat-. George et al. ||Flow cytometry||Counts||Citrate||1500 × g, 15 min 13 000 × g, 2 min||–||Annexin V||CD||CD51, CD144 or CD146||CD45|
|Freyssinet et al. ||Solid phase capture||Prothromb- inase capture||Citrate||1500 × g, 15 min 13 000 × g, 2 min||–||Annexin V, tissue factor||CD62P or GPIba||CD31 or CD62E||CD45|
|Jimenez et al. ||Flow cytometry||Counts||Citrate||200 × g, 10 min 1500 × g, 7 min||–||–||CD41 or CD42b & CD31||CD31+/ CD42– or CD62E||CD45|
|Nomura ||ELISA||Standard PMP||EDTA||1500 × g, 20 min||–||–||GP IX (capture) CD62P, CD40L||–||–|
|Shet et al.||Flow cytometry||Counts||Citrate||13 000 × g, 10 min||100 000 × g, 60 min||Annexin V||CD41a||CD144||CD14 (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.
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