Hi Emanuel, R&D systems have a product called Statusflow Pro which is a stabilised whole blood control enriched with 2% CD34+ cells. The product insert contains instrustions for use with the ISHAGE gating. I tried a vial 6 months ago and it seemed to work well with our ISHAGE template. The CD34+ cells are possibly a little brighter than mobilised PBSC's. Another advantage of this control is it has values for approx 20 different CD antigens. Regards, Glennis White Haematology Laboratory Wellington Hospital New Zealand glennis.white@ccdhb.org.nz ----- Original Message ----- From: Emanuel Raniolo <emanuel.raniolo@imvs.sa.gov.au> To: cyto-inbox Sent: Tuesday, November 27, 2001 1:33 PM Subject: Use of CD34+ internal controls > > Hi all > > Our standard method for CD34 enumeration is a dual platform ISHAGE gated > (wash) method. > In order to establish and validate a single platform (no wash) ISHAGE method > in the laboratory, it would be nice to be able to run an internal control. > > I have used Beckman-Coulter's commercial Stem-Trol control cells for this > purpose, but have been unable to recover the stated assayed concentration. > In fact I my determinations are consistently higher than the allowable +/- > 15% range. > > Apart from some difficulties in gating placement, as these cells display > increased forward and side light scatter characteristics to normal human > haematopoetic precursors, I cannot see why my results are too high (around > 40% above the stated concentration). I practice reverse pipetting, and mix > the fluorospheres adequately prior to dispensing. Single vs dual platform > comparisons for peripheral blood demonstrate a small positive bias in favour > of the single platform method, whereas results are identical for apheresis > product. In addition, the absolute CD45+ count is in general agreement with > the analyser WBC count. > > I would like to ask: > What is the group experience with the use of internal controls for CD34 > enumeration, and in particular with Stem-Trol cells? > Is the stated Stem-Trol assayed concentration and range reflective of common > experience? > Are internal CD34+ controls regularly utilised in routine practice? > What alternative sources are available? > > Thanks > > Emanuel Raniolo > IMVS Laboratories > The Queen Elizabeth Hospital > Department of Haematology > Cell Analysis and Cryopreservation Laboratory > Adelaide, South Australia. > >
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:01:42 EST