Table 3. Competitive engraftment of... | American Society of Hematology

Table 3.

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

Population^3-150	No. of cells injected per bone^3-151		Primary positive grafts^3-152	Primary CB % positive cells^3-153			Expanded positive grafts^3-152	S-F expanded CB % positive cells^3-153
Population^3-150	CD34⁺	CD34⁺CD38⁻	Primary positive grafts^3-152	CD33	CD19	CD34	Expanded positive grafts^3-152	CD33	CD19	CD34
Primary CB	1.5 × 10⁴	8.8 × 10²	20/20	53 ± 8	59 ± 9	62 ± 8
S-F expanded CB	1.5 × 10⁴	Not detected					22/23	59 ± 7	62 ± 8	63 ± 7
Primary CB + S-F
expanded CB	See above		20/22	41 ± 7	48 ± 9	50 ± 9	5/22	18 ± 16	16 ± 14	20 ± 17

Population^3-150	No. of cells injected per bone^3-151		Primary positive grafts^3-152	Primary CB % positive cells^3-153			Expanded positive grafts^3-152	S-F expanded CB % positive cells^3-153
Population^3-150	CD34⁺	CD34⁺CD38⁻	Primary positive grafts^3-152	CD33	CD19	CD34	Expanded positive grafts^3-152	CD33	CD19	CD34
Primary CB	1.5 × 10⁴	8.8 × 10²	20/20	53 ± 8	59 ± 9	62 ± 8
S-F expanded CB	1.5 × 10⁴	Not detected					22/23	59 ± 7	62 ± 8	63 ± 7
Primary CB + S-F
expanded CB	See above		20/22	41 ± 7	48 ± 9	50 ± 9	5/22	18 ± 16	16 ± 14	20 ± 17

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

F3-150

CB CD34⁺ cells were immunomagnetically isolated and expanded for 17 days in S-F cultures supplemented with IL3, IL6, GM-CSF, SCF, and FLT-3L. S-F 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.

F3-151

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

F3-152

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

F3-153

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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