Cryopreservation of rare pediatric red blood cells for support following bone marrow transplant

Author:

Kelly Kathleen1ORCID,Helander Louise23,Hazegh Kelsey1,Stanley Crystal4,Moss Raymond1,Mack Samantha4,Sanders Mary L.3,Gurley Janice3,McKinney Chris56,Dumont Larry J.17ORCID,Annen Kyle8

Affiliation:

1. Vitalant Research Institute Denver Colorado USA

2. ClinImmune Labs/Department of Medicine University of Colorado Aurora Colorado USA

3. Department of Pathology and Laboratory Medicine Children's Hospital Colorado Aurora Colorado USA

4. Vitalant Denver Colorado USA

5. Center for Cancer and Blood Disorders Children's Hospital Colorado Aurora Colorado USA

6. Medical Department of Pediatrics University of Colorado Aurora Colorado USA

7. Department of Pathology and Laboratory Medicine University of Colorado Medical School Aurora Colorado USA

8. Department of Pathology University of Colorado Aurora Colorado USA

Abstract

AbstractBackgroundA 2‐year‐old, 10.8 kg male pediatric patient with X‐linked chronic granulomatous disease (CGD) with McLeod syndrome (MLS) was scheduled for a hematopoietic stem cell transplant (HSCT). Identification of allogenic red blood cells (RBC) for post‐transplant support was unsuccessful prompting the development of a customized method to collect and freeze rare autologous pediatric cells.Study Design and MethodsA protocol was developed for the collection of small volume pediatric whole blood (WB) via peripheral venipuncture with collection into 10 ml syringes containing anticoagulants. Additionally, a closed system RBC glycerolization and deglycerolization instrument was adapted to process small volume, non‐leukoreduced WB. Both collection and WB processes were validated. In total 4 approximately 100 ml autologous units were collected and frozen. Two units were thawed, deglycerolized, and used for clinical transfusion support. To appreciate processing impacts on RBC rigidity, ektacytometry was performed on pre‐processed and post‐deglycerolization samples.ResultsFree hemoglobin (HGB) of validation units after thawing/deglycerolization was <150 mg/dL with an average red cell recovery of 85%. These units also showed little difference between pre‐and post‐processing Lorrca deformability curves or membrane rigidity. Two pediatric units were thawed and deglycerolized for transfusion. Free HGB was 70 mg/dL and 50 mg/dL post‐thaw, and these RBCs had a slight decrease in deformability and increased membrane rigidity.DiscussionCustomized WB collection, glycerolization, freezing, and deglycerolization processes were developed to successfully support a pediatric patient with CGD and MLS after autologous HSCT. Both pediatric units showed increased membrane rigidity post‐deglycerolization which may be a consequence of the CGD and MLS genetic background.

Publisher

Wiley

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