Cells cultured for 7 AFT024 in noncontact culture with FL + SCF + Il-7 produce multilineage engraftment in tertiary fetal sheep recipients.

CB CD34⁺ cells (100 000), cultured in AFT024 noncontact culture for 7 days were injected into 4 fetal sheep. Sixty days after transplantation, 2 fetuses were killed and the human CD45⁺cells pooled and then injected into 3 secondary recipients. These recipients were killed at 60 days, BM pooled, and injected into 3 tertiary recipients. After 60 days, fetuses were killed and the marrow obtained and assessed for multilineage human engraftment by 2-color flow cytometry. Cells were stained with antihuman-CD45, CD20, CD3, CD33, CD34, and glyophorin-A antibodies. Isotype-matched controls are demonstrated in the top right. SSC indicates side scatter; FSC, forward scatter; FL2, fluorochrome 2.