Table 2. Competitive engraftment of... | American Society of Hematology

Table 2.

Competitive engraftment of primary and PMVEC coculture expanded cord blood CD34⁺ cells in triple HLA-mismatched SCID-hu bone model

Population^*	No. of cells injected per bone^†		Primary positive grafts^‡	Primary CB % positive cells^2-153			PMVEC positive grafts^‡	PMVEC expanded CB % positive cells^2-153
Population^*	CD34⁺	CD34⁺CD38⁻	Primary positive grafts^‡	CD33	CD19	CD34	PMVEC positive grafts^‡	CD33	CD19	CD34
Primary CB	1.5 × 10⁴	8.8 × 10²	20/20	53 ± 7	59 ± 9	62 ± 8
PMVEC expanded CB	1.5 × 10⁴	1.8 × 10³					20/21	49 ± 7	72 ± 7	59 ± 7
Primary CB + PMVEC
expanded CB	See above		20/20	40 ± 7	47 ± 8	47 ± 7	2/20	1.4, 42	2, 1	5, 4

Population^*	No. of cells injected per bone^†		Primary positive grafts^‡	Primary CB % positive cells^2-153			PMVEC positive grafts^‡	PMVEC expanded CB % positive cells^2-153
Population^*	CD34⁺	CD34⁺CD38⁻	Primary positive grafts^‡	CD33	CD19	CD34	PMVEC positive grafts^‡	CD33	CD19	CD34
Primary CB	1.5 × 10⁴	8.8 × 10²	20/20	53 ± 7	59 ± 9	62 ± 8
PMVEC expanded CB	1.5 × 10⁴	1.8 × 10³					20/21	49 ± 7	72 ± 7	59 ± 7
Primary CB + PMVEC
expanded CB	See above		20/20	40 ± 7	47 ± 8	47 ± 7	2/20	1.4, 42	2, 1	5, 4

CB indicates cord blood; primary, primary CB; PMVEC and PMVEC expanded, PMVEC coculture expanded CB.

CB CD34⁺ cells were immunomagnetically isolated and cultured for 17 days on pre-established monolayers of porcine microvascular endothelial cells (PMVEC) with IL3, IL6, GM-CSF, SCF, and FLT-3L. PMVEC coculture-expanded CB cells were injected individually or simultaneously with HLA-mismatched primary CB cells into human fetal bone fragment (bearing a third distinct HLA allele) implanted in the SCID mouse (SCID-hu bone assay) to determine their competitive marrow repopulating ability. Grafts were normalized for CD34⁺ cell content prior to injection.

†

Ten weeks after injection, fetal bone implants were removed and analyzed for the presence of primary or PMVEC coculture-expanded CB HLA-bearing cells and CD19, CD33, and CD34 cells in a 5-color flow-cytometric assay. Uninjected grafts were used to determine the level of nonspecific binding of monoclonal antibodies.

‡

Positive grafts indicate number of grafts containing donor-derived multilineage hematopoiesis per total number of injected grafts.

F2-153

Percent positive cells represent donor-derived to total cells of the particular cell lineage present in the bone 10 weeks after injection. Numbers are expressed as mean % ± SEM.

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