A simple filtration system for red blood cell depletion and volume reduction in routine processing of human umbilical cord blood.
OBJECTIVE
Currently, stem cells and other progenitor cells are obtained from human umbilical cord blood (HUCB) using a variety of methods that are designed primarily for red blood cell depletion and volume reduction prior to freezing and storage. Some of these methods are very cumbersome and involve several steps that may result in significant cell loss. Therefore, processes that minimize the loss of haematopoietic stem and progenitor cells (HSPC) remains very critical. In the present study, we describe a simple filtration process for achieving both volume reduction and red blood cell depletion in a 'closed sterile system' with significant recovery of viable HSPC.


METHODS
About 80-100 ml of HUCB were collected into citrate-phosphate-dextrose-adenine 1 anticoagulant. Each HUCB was divided into 25-70-ml aliquots and then either diluted with isotonic saline or filtered without any prior dilution with an experimental Red Cell Volume Reduction System (RCVRS). The HSPCs were recovered by retrograde rinsing of the filter with an isotonic stem cell recovery solution. The viability, colony forming properties, leucocytes and CD34+ cells recoveries were determined.


RESULTS
The mean volume of the HUCB before processing was reduced from 43.9 +/- 7.9 ml to 11.8 +/- 0.7 ml (n = 55) with red blood cell depletion of 85.2 +/- 3.7%. Diluting the HUCB with isotonic saline prior to processing with RCVRS increased the red blood cell depletion to 91.9 +/- 3.0% (n = 7) without any significant loss in viability or cell recovery. The mean viability of the RCVRS-processed HUCB was not significantly different from the control unprocessed blood (96.60 +/- 1.90 vs. 96.63 +/- 2.12%; P > 0.05). The mean recoveries of the CD34+ and the haematopoietic clonogenic progenitor cells with the filter were 83.9 +/- 26.8 (n = 40) and 99.9 +/- 27.9% (n = 35), respectively.


CONCLUSIONS
The present results show that the RCVRS provides a simple and easy-to-use process for obtaining red blood cell depletion and volume reduction of HUCB with good cell viability and recoveries.
Sowemimo-Coker
S O
SO
Pall Medical R&D, Pall Life Sciences, Pall Corporation, Port Washington, NY 11050, USA.
Andrade
F
F
Kim
A
A
Pesci
S
S
D018952:Antigens, CD34 / Q000032:analysis; D002469:Cell Separation / Q000379:methods*; D004912:Erythrocytes / Q000166:cytology; D005312:Fetal Blood / Q000166:cytology*; D005374:Filtration; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D007962:Leukocytes / Q000166:cytology
D018952:Antigens, CD34