Department of Physiology, Kasturba Medical College, MAHE, Manipal, Karnataka, India
*Corresponding Author: Department of Physiology, Kasturba Medical College, MAHE, Manipal, Karnataka, India
Received: February 05, 2020; Published: September 30, 2020
Renal transplantation is the most successful treatment option for chronic renal failure patients. But success of this depends on HLA crossmatching between the renal recipient and donor. In this test, RBCs and platelets are removed and only donor lymphocytes are incubated with recipient’s serum and the complement and percentage of dead lymphocytes is documented. More than 5% of dead cells indicates a positive test and is a contraindication for renal transplantation. For HLA crossmatching, platelets must be removed before lymphocytes mixed with serum. Removal of platelets for crossmatching by traditional method is a very expensive procedure till now as commercially available thrombin is very costly, which increases the financial burden of the patients. Therefore, we tried to develop a cost effective technique to remove platelets by density gradient centrifugation in patients and donors visiting the Nephrology department of Kasturba Hospital, Manipal. After removing plasma and red blood cells, cell suspension in MCT vials is centrifuged @ 1000 rpm for 5 minutes using microcentrifuge. After this, supernatant is discarded and cell pellet is resuspended with buffer and again centrifuged @ 1000 rpm for 5 minutes. This step is repeated 4 or 5 times until the cell suspension is completely cleared off platelets. A total of 96 HLA crossmatching tests done on renal transplant patients and donors visiting Kasturba Hospital, Manipal. Out of this, two cases showed 5% platelets even after 5 times centrifugation. When we centrifuged the cell suspension @ 1000 rpm for 5 minutes, less denser platelets remained in supernatant and more denser lymphocytes sediment to the bottom. When this is repeated 4 or 5 times, all the platelets are removed. This is a very cost effective technique compared to commercial thrombin used earlier, which has reduced the cost of HLA crossmatching.
Keywords: HLA Crossmatch; MCT Vials; CRF
Citation: Murali Adiga. “Cost Effective Technique of Removal of Platelets for HLA Crossmatching by Density Gradient Centrifugation". Acta Scientific Gastrointestinal Disorders 3.10 (2020): 17-19.
Copyright: © 2020 Murali Adiga. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.