Mobilization of Progenitor Cells for Autologous and Allogenei Transplantation
A. Zander1, W. Zeller1, M. Stockschläder1, C. Berger1, W. Krüger1,
J. Grimm1, K. Hummel2, C. Löliger2, P. Kühnl2, H. Kabisch3   

Zander AR et al. Gene Technology,Stem Cell and Leukemia ResearchNato Asi Series H:
Cell Biology, Vol 94,Springer-Verlag, Berlin Heidelberg New York London

1 Dept. Oncology and Hematology, BMT-Unit,
2 Dept. of TransfusionMedicine.
3 Dept. Pediactric Hematology, University Hospital Eppendorf, Hamburg, Germany


The hematopoietic growth factors G-CSF and GM-CSF are able to stimulate granulopoiesis after autologous transplantation of bone marrow or blood stem cells (PBSC) and can also mobilize peripheral blood progenitor cells [1, 2, 3, 4]. Rapid recovery of platelets has been observed following addition of G-CSF mobilized PBSC to autologous bone marrow transplants [3]. The kinetics of engraftment are mainly dependent on the number and qualitiy of progenitor/stem cells in the graft [5, 6, 7, 8]. We have studied the impact of higher doses of G-CSF (Filgrastim) on the yield of CD34 positive cells, CFU-GM and mononucelar cells in leukapheresis products in patients with Non Hodgkin Lymphomas, Hodgkins disease, testicular cancer, AML and neuroblastoma as well as in normal donors for allogeneic progenitor cell transplantation. We report our experience with mobilization of GCSF in two different doses: 10 µg/kg per day versus 24 (2 x 12) µg/kg per day for stem cell collection, separation and transplantation.

Materials and methods

Mobilization: G-CSF (Filgrastim) was administered subcutaneously for 5- 6 days at a daily dose of 10 µg/kg (group A) or 2x12 µg/kg (group B) to 45 patients. Beyond this a total of 18 mobilizations followed by 1 to 4 leukapheresis procedures, was carried out in normal donors. 15 transplants, involving mobilized progenitor cells either alone or in conjunction with bone marrow, were carried out. The characteristics of the autologous patients are shown in table 1.

Table 1. Patient characteristics

15 patients with the following diagnosis: AML: 4, ALL: 5, Multiple Myeloma: 1, Severe Aplastic Anaemia: 3, CML blastic crisis: 1, Myeloproliferative Syndromes 1 received an allogeneic stem cell transplant. The donor-recipient relationships were the following: matched siblings: 8, one mismatch: 2, parent to child: 3, child to parent: 1, matched unrelated donor: 1. Conditioning regimens: without conditioning as a rescue graft for rescue from insufficient bone marrow function: 5; TBICyclophosphamide-VP16: 5; TBI-Cyclophosphamide: 1, Busulfan, Cyclophosphamid +- Etoposid: 4.

Colony forming assay: Colony forming units granulocyte-monocyte (CFU-GM) were assayed in Iscove's methylcellulose. Colonies were enumerated after 14 days of culture in 37° C, 5 % CO2 and 100 % humidity by using an inverted phase microscope (9).

Flow cytometric analysis: Surface antigen expression was evaluated using the monoclonal antibody HPCA-2, directly conjugated to FITC (Becton-Dickinson). Analysis was performed on a FACScan (Becton-Dickinson) using Lysis II research software [5,10,11].

Statistics: Data are shown as median (range), statistical significance was determined by the Student's t-test.


Autologous: The percentage of CD34+ mononuclear cells (MNC) in the first leukapheresis product increased significantly (p=0.024) from 0.68 (0.03-1.96) (group A) to 1.23 (0.4-4.28) (group B) (Fig. 1 ).

Fig.1 Leukapheresis products 10µg vs. 2x12µg G-CSF

Statistically significant differences were also found with respect to total CD34+ MNC per leukapheresis product (LP1): 10.08x10high 7 (0.4-44.0) (group A) versus 42.51 x 10 high 7 (3.46-86.3) (group B) (p=0.012) (Fig. 2, Tabl. 2).

Fig. 2 Leukapheresis products 10µg vs. 2x12µg G-CSF

Table 2. Graft Data

CFU-GM increased from 37.05 (1.6- 95.1) x 105 (group A) to 106.5 (8.7- 390.5) (group B) (p = 0.004). These results confirm, that higher doses of G-CSF can significantly increase the number of CD34 positive cells and CFU-GM in leukapheresis products.

Allogeneic: An average of 3 leukapheresis (range 1 to 4) were carried out, which yielded 13 x 10 high8 (range 2.9 -29) mononuclear cells/kg BW of the recipient, 5 x 10 high 6 CD34 positive cells/kg BW of recipient (range 1.2- 39), CFC 3 x 10 high 5 /kg BW (0,88 60,4). The mobilization with G-CSF was generally well tolerated with the exception of moderate bone pain in 2/3 of the donors.

Engraftment in 9 evaluable patients revealed neutrophil engraftment of more than 0,5/nl by day 14 (range 9 -22), neutrophils more than 1.0/nl day 15 (range 10- 24), thrombocytes more than 20/nl day 16 (range 8 -32), more than 50/nl 24 (range 13 to more than 100 days).

GvH grade 0 -1: 3 patients, grade 2: 4 patients, grade 4: 2 patients and moderate chronic GvH in 1 patient. Total of 12 of the 15 patients are alive in continuous complete remission, 3 have died of GvH and infection (aspergillus 1, candida crusei 1 ).


We could show that the mobilization with a higher than usual dosis of G-CSF is well tolerated in normal donors as well as in patients and yields grafts which allow engraftment after myeloablative therapy. GvH occured in 2/3 of evaluable patients and led in conjunction with infection to death in 3 patients. 1 patient received an allogeneic graft from a matched unrelated donor, showed engraftment but unfortunately died of a preexisting fungus disease.

Based on our experience in the autologous setting, where we found statistically significant differences in the number of CD34+ cells and CFU-GM in patients mobilized with 10 µg/kg G-CSF compared with 24 µg/kg and based on the observation that the side effects in both dose levels are comparable, we would recommend 24 µg/kg BW G-CSF per day, given in 2 divided doses or continuously intravenous infusion for mobilization of normal donors.


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