American Society of Hematology

Fig. 3.

$Fig. 3. Two different B220+ lymphocytes derived from the CD34+ EB cell fraction. (A) Morphological characterization by Wright-Giemsa staining (Objective ×100 Oil). Type II LAK (LAK34a-2) cells (left), type I LAK (LAK34a-3) cells (center), and EB34-pre-B cells (right) were spun on slides and stained with Wright-Giemsa. Large granules are present in both small LAK34a-2 cells and larger LAK34a-3 cells, but not in EB34-pre-B cells. (B) Phenotypic analysis of CD34+ EB cell– derived LAK cells. Type II LAK cells (LAK34a-2, upper panels) and type I LAK cells (LAK34a-3, lower panels) were stained with PE-conjugated anti-CD3, Mac-1, Sca-1, and B220 monoclonal antibodies. Positive cells for CD3, Mac-1, Sca-1, and B220 (region ⊢⊣) are 0.1%, 23.0%, 40.5%, and 99.1%, respectively, in LAK34a-2, and 0.2%, 62.9%, 98.9%, and 98.5%, respectively, in LAK34a-3. Note the difference in the Sca-1 pattern between LAK34a-2 and LAK34a-3. Results of the corresponding isotype control are indicated in gray. (C) Phenotypic analysis of EB34-pre-B cells. Adherent EB34-pre-B cells were collected from the OP9 stroma layer, and stained with PE-conjugated anti-CD43, B220, BP-1, and CD19 monoclonal antibodies. Positive cells for CD43, B220, BP-1, and CD19 (region ⊢⊣) are 99.8%, 66.5%, 0%, and 99.5%, respectively. Background staining is indicated in gray.$

Two different B220⁺ lymphocytes derived from the CD34⁺ EB cell fraction. (A) Morphological characterization by Wright-Giemsa staining (Objective ×100 Oil). Type II LAK (LAK34a-2) cells (left), type I LAK (LAK34a-3) cells (center), and EB34-pre-B cells (right) were spun on slides and stained with Wright-Giemsa. Large granules are present in both small LAK34a-2 cells and larger LAK34a-3 cells, but not in EB34-pre-B cells. (B) Phenotypic analysis of CD34⁺ EB cell– derived LAK cells. Type II LAK cells (LAK34a-2, upper panels) and type I LAK cells (LAK34a-3, lower panels) were stained with PE-conjugated anti-CD3, Mac-1, Sca-1, and B220 monoclonal antibodies. Positive cells for CD3, Mac-1, Sca-1, and B220 (region ⊢⊣) are 0.1%, 23.0%, 40.5%, and 99.1%, respectively, in LAK34a-2, and 0.2%, 62.9%, 98.9%, and 98.5%, respectively, in LAK34a-3. Note the difference in the Sca-1 pattern between LAK34a-2 and LAK34a-3. Results of the corresponding isotype control are indicated in gray. (C) Phenotypic analysis of EB34-pre-B cells. Adherent EB34-pre-B cells were collected from the OP9 stroma layer, and stained with PE-conjugated anti-CD43, B220, BP-1, and CD19 monoclonal antibodies. Positive cells for CD43, B220, BP-1, and CD19 (region ⊢⊣) are 99.8%, 66.5%, 0%, and 99.5%, respectively. Background staining is indicated in gray.

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