Recovery and functional activity of mononuclear bone marrow and peripheral blood cells after different cell isolation protocols used in clinical trials for cell therapy after acute myocardial infarction.
OBJECTIVE
Clinical trials showed contradictory results in functional recovery after intracoronary infusion of autologous mononuclear (bone marrow) cells in patients with acute myocardial infarction. A recent study suggests that this might be related to the isolation protocol used. In The Netherlands, a comparable randomised multicentre trial (HEBE) was designed. To validate the isolation method of bone marrow and peripheral blood-derived mononuclear cells, we compared our processing protocol with methods comparable to the ASTAMI (no beneficial effect) and the REPAIR-AMI study (beneficial effect).


RESULTS
The effect of several factors (density gradient, washing buffer and centrifugation speed) has been studied on recovery and function (migration and clonogenic capacity) of mononuclear cells. Significantly lower cell recoveries were found at a centrifugation speed of 250 g, compared to 600 or 800 g, respectively. Furthermore, washing buffer without supplemented human serum albumin and heparin resulted in significantly lower cell recovery and functional impairment as measured by clonogenic capacity.


CONCLUSIONS
The results of our study justify the cell-processing protocol as applied in the HEBE trial (600 g, human serum albumin supplemented washing buffer). This protocol results in viable and functional cells of which the quantity and quality is at least comparable to a successful study like the REPAIR-AMI.
van Beem
Rachel T
RT
Department of Experimental Immunohematology, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
Hirsch
Alexander
A
Lommerse
Ingrid M
IM
Zwaginga
Jaap Jan
JJ
Noort
Willy A
WA
Biemond
Bart J
BJ
Piek
Jan J
JJ
van der Schoot
C Ellen
CE
Voermans
Carlijn
C
D000925:Anticoagulants / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects; D016026:Bone Marrow Transplantation; D002021:Buffers; D006348:Cardiac Surgical Procedures; D002465:Cell Movement; D002469:Cell Separation / Q000379:methods* / Q000592:standards*; D002498:Centrifugation / Q000379:methods; D002985:Clinical Protocols; D002986:Clinical Trials as Topic; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000187:drug effects; D009203:Myocardial Infarction / Q000601:surgery*; D012709:Serum Albumin / Q000494:pharmacology; D033581:Stem Cell Transplantation; D013249:Sternum / Q000166:cytology*
D000925:Anticoagulants; D002021:Buffers; D012709:Serum Albumin; D006493:Heparin