Table 5. Competitive engraftment of... | American Society of Hematology

Table 5.

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

Population^5-150	No. of cells injected per bone^5-151		ABM positive grafts^5-152	ABM-PMVEC % positive cells^5-153			CB positive grafts^5-152	CB-PMVEC % positive cells^5-153
Population^5-150	CD34⁺	CD34⁺CD38⁻	ABM positive grafts^5-152	CD33	CD19	CD34	CB positive grafts^5-152	CD33	CD19	CD34
ABM-PMVEC	1 × 10⁵	3.2 × 10³	10/10	26 ± 9	39 ± 12	29 ± 11
CB-PMVEC	1 × 10⁵	1.7 × 10⁴					9/9	42 ± 10	56 ± 12	40 ± 9
ABM-PMVEC + CB-PMVEC	See above		0/6	0	0	0	6/6	0.3 ± 0.1	17 ± 7	5 ± 2

Population^5-150	No. of cells injected per bone^5-151		ABM positive grafts^5-152	ABM-PMVEC % positive cells^5-153			CB positive grafts^5-152	CB-PMVEC % positive cells^5-153
Population^5-150	CD34⁺	CD34⁺CD38⁻	ABM positive grafts^5-152	CD33	CD19	CD34	CB positive grafts^5-152	CD33	CD19	CD34
ABM-PMVEC	1 × 10⁵	3.2 × 10³	10/10	26 ± 9	39 ± 12	29 ± 11
CB-PMVEC	1 × 10⁵	1.7 × 10⁴					9/9	42 ± 10	56 ± 12	40 ± 9
ABM-PMVEC + CB-PMVEC	See above		0/6	0	0	0	6/6	0.3 ± 0.1	17 ± 7	5 ± 2

CB indicates cord blood; ABM, adult bone marrow; ABM-PMVEC, PMVEC coculture expanded ABM, CB-PMVEC, PMVEC coculture expanded CB.

F5-150

CB and ABM 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-expanded CB and ABM 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. CD34⁺ cells were reisolated prior to injection.

F5-151

Ten weeks after injection, fetal bone implants were removed and analyzed for the presence of CB and ABM 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.

F5-152

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

F5-153

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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