Table 4. Competitive engraftment of... | American Society of Hematology

Table 4.

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

Population^4-150	No. of cells injected per bone^4-151		S-F positive grafts^‡	S-F expanded CB % positive cells^4-153			PMVEC positive grafts^‡	PMVEC expanded CB % positive cells^4-153
Population^4-150	CD34⁺	CD34⁺CD38⁻	S-F positive grafts^‡	CD33	CD19	CD34	PMVEC positive grafts^‡	CD33	CD19	CD34
S-F expanded CB	1.5 × 10⁴	not detected	22/23	59 ± 7	62 ± 7	63 ± 7
PMVEC expanded CB	1.5 × 10⁴	1.8 × 10³					20/21	49 ± 7	72 ± 7	59 ± 7
S-F expanded CB + PMVEC
expanded CB	See above		18/18	33 ± 4	46 ± 6	42 ± 5	11/18	29 ± 6	24 ± 6	26 ± 6

Population^4-150	No. of cells injected per bone^4-151		S-F positive grafts^‡	S-F expanded CB % positive cells^4-153			PMVEC positive grafts^‡	PMVEC expanded CB % positive cells^4-153
Population^4-150	CD34⁺	CD34⁺CD38⁻	S-F positive grafts^‡	CD33	CD19	CD34	PMVEC positive grafts^‡	CD33	CD19	CD34
S-F expanded CB	1.5 × 10⁴	not detected	22/23	59 ± 7	62 ± 7	63 ± 7
PMVEC expanded CB	1.5 × 10⁴	1.8 × 10³					20/21	49 ± 7	72 ± 7	59 ± 7
S-F expanded CB + PMVEC
expanded CB	See above		18/18	33 ± 4	46 ± 6	42 ± 5	11/18	29 ± 6	24 ± 6	26 ± 6

CB indicates cord blood; S-F and S-F expanded, stroma-free expanded, PMVEC and PMVEC expanded, PMVEC coculture expanded CB.

F4-150

CB CD34⁺ cells were immunomagnetically isolated and expanded for 17 days in cultures with or without pre-established PMVEC monolayers. Cultures were supplemented with IL3, IL6, GM-CSF, SCF, and FLT-3L. HLA-mismatched PMVEC coculture and S-F expanded CB cells were injected individually or simultaneously 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 before injection.

F4-151

Ten weeks after injection, fetal bone implants were removed and analyzed for the presence of PMVEC coculture or S-F 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.

F4-153

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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