Isolation and storage of peripheral blood hematopoietic stem cells for
autotransplantation into children with cancer
Y Takaue, T Watanabe, Y Kawano, T Koyama, T Abe, T Suzue, J Satoh, T Shimokawa, T Ninomiya and M Kosaka
Department of Pediatrics, University of Tokushima, Japan.
Peripheral blood stem cells (PBSC) were collected for autotransplantation
by a total of 46 continuous-flow leukaphereses in 17 children with various
types of cancer in whom the stem-cell pool had been expanded by
chemotherapy. As the cells collected by leukapheresis were contaminated
with many visible cell clumps, platelets, and erythrocytes, they were
separated from the platelet-rich plasma (PRP) by slow-speed centrifugation
and fractionated on a discontinuous gradient of Percoll. All the
hematopoietic progenitors (CFU-GM, CFU- GEMM) in the starting samples were
recovered at the interface of 40% and 60% Percoll solutions largely free of
other cellular components and with a substantial reduction in volume. The
separation and freezing procedures could be completed within three hours
after obtaining cells by leukapheresis. After their fractionation and
storage, these PBSC were shown to be able to reconstitute normal
hematopoiesis in ten children with poor prognosis leukemia or neuroblastoma
for whom no HLA- compatible marrow donors were available and who had been
subjected to marrow-ablative therapy. This separation procedure is simple,
efficient, and readily available and can be used for children as a routine
procedure for PBSC autotransplantation.
Volume 74,
Issue 4,
pp. 1245-1251,
09/01/1989
Copyright © 1989 by The American Society of Hematology
