Quantification of Nucleated Cells, CD34-Positive Cells and CFU-GM Colonies in Single Bone Marrow Samples and Bone Marrow Harvests Derived from Healthy Children

Abstract

Little is known regarding bone marrow (BM) cellularity, CD34+ fraction, and CFU-GM colony formation in relation to age and whether healthy children require a reference range distinct from healthy adults. We therefore analyzed a series of single BM aspirates from 45 healthy children who were evaluated as potential BM donors. Thirty-three of these children subsequently donated BM. We quantified the nucleated cell count, fraction of CD34+ cells, and number of CFU-GM colonies in single aspirates and BM harvests. Single aspirates displayed a mean nucleated cell count of 31.3 × 10⁶ cells/mL, a mean fraction of 1.17% CD34+ cells, and a mean colony forming potential of 66.6 CFU-GM/10⁵ cells. Harvests yielded the same number of nucleated cells but increased numbers of CD34+ cells and CFU-GM compared with single aspirates. The mean nucleated cell count in BM harvests was 31.1 × 10⁶ /mL with a mean fraction of 1.95% CD34+ cells and a mean of 112.4 CFU-GM colonies/10⁵ cells. The concentration of nucleated cells was elevated compared with reported adult counts, while CD34+ percentage and CFU-GM counts were similar. In this series of healthy children, the fraction of CD34+ cells, CFU-GM colonies, and nucleated cells decreased with age. We did not identify gender specific differences. To our knowledge, this represents the first comprehensive study of CD34+ cell fraction, CFU-GM counts, and nucleated cell numbers in the BM of healthy children. The findings provide valuable information for practical use for BM transplantation and contribute to the understanding of hematopoiesis from birth to adulthood.

Keywords :

• bone marrow aspirates versus BM harvests
• bone marrow nucleated cells
• CD34-positive cells
• CFU-GM colonies in healthy children

ABBREVIATIONS
BM	=	Bone marrow
CD	=	Cluster of differentiation
CFU-GM	=	Colony-forming unit granulocyte/macrophage
EDTA	=	Ethylene diamine tetraacetate
FITC	=	Fluorescein isothiocyanate
G-CSF	=	Granulocyte colony-stimulating factor
ISHAGE	=	International Society for Hematotherapy and Graft Engineering
PE	=	Phycoerithrin
SD	=	Standard deviation
SE	=	Standard error