Table 4. Phenotype of Cells Generated by... | American Society of Hematology

Table 4.

Phenotype of Cells Generated by Human CD34⁺ or CD34⁺CD38^low Cells Sorted From the Marrow of SCID or NOD-SCID Mice Transplanted With Human CB-MNC or CD34⁺ Cells and Cultured With MS-5 Stromal Cells

Experiment No.	Cells Injected	Mice	Time (wk)	Sorted Cells	Time in Culture (wk)	24 Wells^4-150		Cultures at Limiting Dilutions
						CD11b⁺ or CD15⁺	CD19⁺	Cells/Well	No. of Wells	Positive/Total Wells Analyzed^4-151
										CD19⁺	CD19⁺ CD15⁻	CD19⁻ CD15⁺
										CD15⁺/CD11b⁺
1	MNC	SCID	4	34⁺38^low	4	59%	5%	1	180	5/80	4/80	10/80
2	MNC	SCID	7	34⁺38^low	7	25%	10%	ND	ND	ND	ND	ND
3	MNC	NOD-SCID	4	34⁺38^low	4	ND	ND	1	240	3/13	2/13	8/13
								10	60	8/18	4/18	5/18
4	CD34⁺	NOD-SCID	7	34⁺38^low	4	39%	5%	1	60	0/12	9/12	3/12
								10	60	13/18	3/18	2/18
								50	60	10/10	0/10	0/10
5	CD34⁺	NOD-SCID	5	34⁺	7	25%	8%	ND	ND	ND	ND	ND

Experiment No.	Cells Injected	Mice	Time (wk)	Sorted Cells	Time in Culture (wk)	24 Wells^4-150		Cultures at Limiting Dilutions
						CD11b⁺ or CD15⁺	CD19⁺	Cells/Well	No. of Wells	Positive/Total Wells Analyzed^4-151
										CD19⁺	CD19⁺ CD15⁻	CD19⁻ CD15⁺
										CD15⁺/CD11b⁺
1	MNC	SCID	4	34⁺38^low	4	59%	5%	1	180	5/80	4/80	10/80
2	MNC	SCID	7	34⁺38^low	7	25%	10%	ND	ND	ND	ND	ND
3	MNC	NOD-SCID	4	34⁺38^low	4	ND	ND	1	240	3/13	2/13	8/13
								10	60	8/18	4/18	5/18
4	CD34⁺	NOD-SCID	7	34⁺38^low	4	39%	5%	1	60	0/12	9/12	3/12
								10	60	13/18	3/18	2/18
								50	60	10/10	0/10	0/10
5	CD34⁺	NOD-SCID	5	34⁺	7	25%	8%	ND	ND	ND	ND	ND

Cord blood cells (20 × 10⁶ MNC or 25 to 50 × 10⁴ CD34⁺ cells) were injected intravenously to either SCID (experiments no. 1 and 2) or NOD-SCID (experiments 3 through 5) mice. After 4 to 7 weeks, mice were killed and marrow cells were analyzed for the presence of human cells (see Table 3). Human CD34⁺CD38^low cells were sorted by FACS and used to initiate long-term cocultures on MS-5 cells as described in the Materials and Methods. Cells were incubated either in 24 wells (2,500 to 5,000 CD34⁺CD38^low cells/well) or at limiting dilutions in 96-well plates. After 5 to 7 weeks in culture, the phenotype of cultured cells was analyzed by labeling with antibodies directed against human CD19 and CD15 (or CD11b).

Abbreviation: ND, not done.

F4-150

Percentages represent the proportion of nucleated cells of the indicated phenotype.

F4-151

Total wells analyzed represent the number of wells in which enough cells were detectable to allow reliable phenotypic analysis. Positive wells represent the number of these analyzed wells that contain cells of the indicated phenotype.

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