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. 2013 Jan;52(1):70-7.

Leukapheresis protocol for nonhuman primates weighing less than 10 kg

Vimukthi Pathiraja  1 , Abraham J Matar, Ashley Gusha, Christene A Huang, Raimon Duran-Struuck
Affiliations

Leukapheresis protocol for nonhuman primates weighing less than 10 kg

Vimukthi Pathiraja et al. J Am Assoc Lab Anim Sci. 2013 Jan.

Abstract

Leukapheresis is a common procedure for hematopoietic cell transplantation in adults. The main challenge in applying this procedure to human infants and small monkeys is the large extracorporeal blood volume (165 mL on average) necessary for priming the apheresis machine. This volume represents greater than 50% of the total circulating blood volume of a human neonate or small monkey. In this report, we document a safe leukapheresis protocol developed for rhesus macaques (3.9 to 8.7 kg). To avoid sensitizing donor animals undergoing leukapheresis to third-party blood products, autologous blood collected during the weeks prior to leukapheresis was used to volume-expand the same donor while priming the machine with saline on the day of leukapheresis. During the procedures, blood pressure was controlled by monitoring the inlet volume, and critical-care support was provided by the anesthesia team. Electrolytes and hemogram parameters were monitored intermittently. Overall, our research subjects underwent effective 4- to 6-h leukapheresis. A total of 9 leukapheresis procedures were performed, which yielded 1 × 10(9) to 6 × 10(9) peripheral blood mononuclear cells containing 1.1 to 5.1 × 10(6) CD34(+) cells (assessed in 4 of 9 macaques) in a volume of 30 to 85 mL. All macaques showed decreases in Hct and platelet counts. In summary, we report a successful modified leukapheresis procedure that can be performed safely in small animals without modification of the leukapheresis machine or associated cell-collection kits.

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Figures

Figure 1.
Figure 1.
WBC counts in macaques that received (top left) 10 μg/kg GCSF, (top right) 100 μg GCSF, or (bottom) 100 μg/kg GCSF + 1 mg/kg AMD3465. Macaques in all groups exhibited an increase or maintenance of WBC counts during leukapheresis. The regimens using 100-μg/kg GCSF led to markedly increased cell counts after mobilization, which were maintained throughout leukapheresis.
Figure 2.
Figure 2.
Hct in macaques that received (top left) 10 μg/kg GCSF, (top right) 100 μg GCSF, or (bottom) 100 μg/kg GCSF + 1 mg/kg AMD3465. Macaques in all groups exhibited mild to moderate decreases in hematocrit. Macaque 0509 (10 μg/kg GCSF) died of hemorrhage.
Figure 3.
Figure 3.
Platelet counts in macaques that received (top left) 10 μg/kg GCSF, (top right) 100 μg GCSF, or (bottom) 100 μg/kg GCSF + 1 mg/kg AMD3465. Macaques in all groups exhibited moderate decreases in platelet counts. With the exception of animal 0509, all macaques maintained platelet counts above 100,000 per microliter.
Figure 4.
Figure 4.
Total protein in leukapheresed macaques. Time 0 is preleukapheresis. Total protein was tested throughout the procedure to tailor albumin supplementation.
Figure 5.
Figure 5.
Total serum calcium in leukapheresed. Time 0 is preleukapheresis. Calcium was tested throughout the procedure while Ca gluconate was given.
Figure 6.
Figure 6.
Blood pressure in leukapheresed macaques. The top 3 panels represent macaques given 10 μg/kg GCSF. The animals shown in the middle 2 panels received 100 μg/kg GCSF. All 4 panels on the bottom depict macaques that received 100 μg/kg GCSF + 1 mg/kg AMD3465.
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