Concentration of bone marrow mononuclear cells using a programmable blood cell separator

Abstract

Bone marrow was collected from adult patients with various solid tumors who consented to participate in a study of myelo-ablative chemotherapy followed by autologous bone marrow rescue. Twenty marrow suspensions were processed by using standard Procedure 3 (PRO-3) for lymphocytapheresis without modification. A modified Procedure 1 (M-PRO-1) for plateletpheresis was employed for processing 34 marrow suspensions. For PRO-3, mononuclear cell (MNC) recovery was 68 +/- 22% of the starting marrow suspension (baseline), in a concentrate volume of 234 +/- 53 ml. MNC represented 59 +/- 27% of the total WBC count of the concentrate. The residual volume of RBC was 49 +/- 47 ml. For M-PRO-1, MNC recovery was 63 +/- 22% of the baseline in a concentrate volume of 200 +/- 8 ml. MNC comprised 94 +/- 7% of the total WBC count of the concentrate. RBC contamination was 7 +/- 3 ml. Hematopoietic recovery, defined as the post-transplant days when a sustained granulocyte count of 500/microL and a platelet count of 50,000/microL were achieved, was similar in the two groups and comparable to other reports utilizing other methods and equipment for bone marrow concentration. Personnel time was significantly reduced compared to other procedures for bone marrow concentration due to increased automation. Although there was no significant difference in MNC recovery between the two groups (P greater than 0.5), M-PRO-1 was clearly superior to PRO-3 because of the consistently high degree of purity of the MNC in the concentrate and minimal RBC contamination.(ABSTRACT TRUNCATED AT 250 WORDS)

A flow cytometric method for detection and enumeration of low‐level, residual red blood cells in platelets and mononuclear cell products

“…There may be situations, however, where the assumptions in automated hematology analyzer algorithms are violated as shown here with MNC collections. As discussed by Orlina and colleagues, 4 secondary processing of marrow MNCs is necessary to reduce rRBCs before transfusion in ABO‐mismatched situations 4 . MNC collections from peripheral blood may also require removal of rRBCs before subsequent processing such as freezing or in vitro culturing and cell differentiation steps.…”

“…Limits of detection and linear ranges for these methods were generally not reported by the investigators 5,6 . In the evaluation of a new PLT apheresis device, Fournel and colleagues 4 used a microscopic method with a Neubauer chamber with a theoretical limit of detection of 1 RBC per 12.5 µL. Linearity and sensitivity was not reported.…”