Korean J Blood Transfus.  1997 Dec;8(2):125-135.

Cryopreservation and Characterization of Umbilical Cord Blood

• Nahm CH
• Lee HB
• Kim CS
• Kim SK
• Im MW
• Kim EY
• Koo MS
• Choi JW
• Kim JJ
• Pai SH

Abstract

BACKGROUND
Umbilical cord blood (UCB) has been increasingly utilized for reconstituting hematopoiesis in a variety of congenital disorders and hematologic malignancies. In this report, we evaluated the maximum collection volume, the efficacy of red cell depletion, cell viability and phenotypic analysis before and after cryopreservation.
METHODS
Forty units of UCB (17 from NSVD and 23 from cesarean section) were collected into blood donation bag with ACD-A. Red cells were depleted using 3% gelatin sedimentation or buffy coat separation. UCB units were cultured in methylcellulose media, and phenotypic analyses were performed with monoclonal antibodies for CD34, HLA-DR, CD38, CD13, CD33, c-kit, CD45/CD3, CD45/CD19, CD3/CD4, CD3/CD8.
RESULTS
The mean volume of one unit of UCB was 109.2 +/- 32.5 mL and one unit contained 1.20 +/- 0.51x19(9) nucleated cells. Cell counts after red cell depletion by 3% gelatin sedimentation or buffy coat separation revealed recovery rates of 77.5 +/- 14.9% and 64.5 +/- 7.4%, respectively. Cell viabilities and the number of colony forming units - granulocyte and monocyte from fresh and cryopreserved UCB were were 98.1 +/- 1.6%, 88.7 +/- 4.8%, and 6.48 +/- 2.14x10(5), 7.35 +/- 0.65x10(5). The mean percentage of CD34+ cells was 1.02 +/- 1.6% and those of CD34+/HLA-DR+, CD34+/CD38+, CD34+/CD13+, CD34+/CD33+, CD34+/c-kit+ cells were 68.6%, 58.0%, 5.6%, 46.8%, and 29.8%, respectively. Lymphocyte subsets were composed of CD45+/CD3+ (59.0%), CD45+/CD19+ (13.8%), CD3+/CD4+ (42.7%), and CD3+/CD8+ cells (17.1%). There was no significant difference in phenotypic characteristics between fresh and cryopreserved UCB.
CONCLUSION
We established the protocols for UCB collection, red cell depletion, cryopreservation, culture and phenotypic analyses. These results would be very useful for future UCB transplantation and preservation/storage.

Keyword

umbilical cord blood; cryopreservation; characterization

MeSH Terms

Antibodies, Monoclonal
Blood Donors
Cell Count
Cell Survival
Congenital, Hereditary, and Neonatal Diseases and Abnormalities
Cryopreservation*
Fetal Blood*
Gelatin
Granulocytes
Hematologic Neoplasms
Hematopoiesis
HLA-DR Antigens
Humans
Lymphocyte Subsets
Methylcellulose
Monocytes
Stem Cells
Umbilical Cord*
Antibodies, Monoclonal
Gelatin
HLA-DR Antigens
Methylcellulose