Table 3. Engraftment of freshly thawed... | American Society of Hematology

Table 3.

Engraftment of freshly thawed and ex vivo expanded umbilical cord blood CD34⁺ cells in different culture conditions

	Day 0		Day 7						Day 14
	Day 0		FS7-SNC^3-152		FT-SNC^3-152		Stroma free^3-153		FS7-SNC^3-152		FT-SNC^3-152		Stroma free^3-153
Cell dose × 10³^3-155
25	5/6^3-150	2.4 ± 1.0^3-151	6/7^3-151	6.0 ± 3.5^3-151	2/4^3-150	9.5 ± 5.7^3-151	1/6^3-150	0.1 ± 0.1^3-151	4/6^3-150	3.3 ± 1.6^3-151	3/4^3-150	3.9 ± 2.5^3-151	2/3^3-150	0.7 ± 0.2^3-151
50	7/7	4.7 ± 1.3	8/8	5.6 ± 1.8	7/8	8.7 ± 3.5	4/5	2.4 ± 1.9	4/6	6.3 ± 3.6	6/6	8.1 ± 3.1	3/3	4.3 ± 1.3
75	4/6	9.0 ± 4.2	4/4	7.1 ± 5.6	5/6	7.8 ± 4.0	3/4	3.5 ± 2.2	5/5	20.4 ± 10.2	5/5	7.0 ± 2.3	4/4	4.5 ± 2.8
100	7/7	13.1 ± 3.4	6/6	15.8 ± 2.9	3/3	9.7 ± 5.5	2/4	14.0 ± 10.4	4/4	11.6 ± 10.1	6/6	15.5 ± 5.0	2/2	9.5 ± 5.5
SRC/10⁵cells^3-154		3.8 (2.2-6.5)		6.5 (3.4-12.5)		3.2 (1.8-5.7)		1.4 (0.7-2.7)		3.6 (2-6.4)		7.2 (3.2-16.6)		6.1 (2.4-15.6)
CD14^#		17.5 ± 4.4		24.1 ± 4.3		16 ± 4.6		5.0 ± 1.0		19.8 ± 3.6		12 ± 0.4		15.3 ± 2.6
CD19		48.8 ± 7.7		31.3 ± 5.3		47.7 ± 5.2		71 ± 2.0		33.3 ± 6.9		54.3 ± 1.5		46 ± 3.1
CD33		42.0 ± 7.0		55.9 ± 5.8		40.3 ± 2.3		25 ± 9.0		50.2 ± 7.8		31.5 ± 3		40 ± 2.6
CD34		13.8 ± 1.0		10.6 ± 1.6		15.3 ± 2.7		29.5 ± 4.5		12.3 ± 2.9		15.8 ± 1.8		13 ± 2.1

	Day 0		Day 7						Day 14
	Day 0		FS7-SNC^3-152		FT-SNC^3-152		Stroma free^3-153		FS7-SNC^3-152		FT-SNC^3-152		Stroma free^3-153
Cell dose × 10³^3-155
25	5/6^3-150	2.4 ± 1.0^3-151	6/7^3-151	6.0 ± 3.5^3-151	2/4^3-150	9.5 ± 5.7^3-151	1/6^3-150	0.1 ± 0.1^3-151	4/6^3-150	3.3 ± 1.6^3-151	3/4^3-150	3.9 ± 2.5^3-151	2/3^3-150	0.7 ± 0.2^3-151
50	7/7	4.7 ± 1.3	8/8	5.6 ± 1.8	7/8	8.7 ± 3.5	4/5	2.4 ± 1.9	4/6	6.3 ± 3.6	6/6	8.1 ± 3.1	3/3	4.3 ± 1.3
75	4/6	9.0 ± 4.2	4/4	7.1 ± 5.6	5/6	7.8 ± 4.0	3/4	3.5 ± 2.2	5/5	20.4 ± 10.2	5/5	7.0 ± 2.3	4/4	4.5 ± 2.8
100	7/7	13.1 ± 3.4	6/6	15.8 ± 2.9	3/3	9.7 ± 5.5	2/4	14.0 ± 10.4	4/4	11.6 ± 10.1	6/6	15.5 ± 5.0	2/2	9.5 ± 5.5
SRC/10⁵cells^3-154		3.8 (2.2-6.5)		6.5 (3.4-12.5)		3.2 (1.8-5.7)		1.4 (0.7-2.7)		3.6 (2-6.4)		7.2 (3.2-16.6)		6.1 (2.4-15.6)
CD14^#		17.5 ± 4.4		24.1 ± 4.3		16 ± 4.6		5.0 ± 1.0		19.8 ± 3.6		12 ± 0.4		15.3 ± 2.6
CD19		48.8 ± 7.7		31.3 ± 5.3		47.7 ± 5.2		71 ± 2.0		33.3 ± 6.9		54.3 ± 1.5		46 ± 3.1
CD33		42.0 ± 7.0		55.9 ± 5.8		40.3 ± 2.3		25 ± 9.0		50.2 ± 7.8		31.5 ± 3		40 ± 2.6
CD34		13.8 ± 1.0		10.6 ± 1.6		15.3 ± 2.7		29.5 ± 4.5		12.3 ± 2.9		15.8 ± 1.8		13 ± 2.1

UCB CD34⁺ cells were cultured in AFT024-noncontact culture or stroma-free culture as described in “Materials and methods.” After 1 and 2 weeks, cell were harvested and transplanted in limiting dilutions in sublethally irradiated NOD-SCID mice as described in “Materials and methods.” After 6 weeks, animals were killed and evaluated for human engraftment.

See Tables 1 and 2 for abbreviations.

F3-150

Number of animals engrafted/number of animals that were transplanted.

F3-151

Mean ± SEM percent human CD45⁺ cells in mouse marrow.

F3-152

AFT024-noncontact culture with 10 ng/mL FL, 10 ng/mL SCF, and 10 ng/mL IL-7 or 10 ng/mL FL and 10 ng/mL TPO.

F3-153

Stroma-free culture contained G-CSF, MIP 1-α, IL-6, MCP-1, VEGF, IL-8, N-desulfated, O-sulfated heparin, Tpo, FL, SCF, and IL-7 engraftment.

F3-155

Determination of SCID repopulating cell (SRC) frequency (see “Materials and methods”).

F3-154

Cells were further analyzed by flow cytometry to demonstrate multilineage differentiation.

#Engraftment data and SRC frequency are derived from 2 independent experiments with 2 to 4 mice being analyzed at each dose and time point. Multilineage potential is the mean ± SEM of 2 to 4 mice.

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