Figure 4.
Figure 4. Cell development in short-term culture assays. ALDHneg CD34+, ALDHbr CD34+, and ALDHbr CD34neg cells were purified from linneg SSClo UCB, as depicted in Figure 1. Two hundred to 1000 purified cells were cultured on STO fibroblasts in the presence of IL-3, IL-7, and IL-15 (n = 10). Cultures initiated with ALDHneg CD34+ cells contained higher percentages of CD56+ lymphoid progeny than did the ALDHbr CD34+ cells (compare panels A and B; see Table 4). Similarly, the ALDHneg CD34+ cell fraction yielded lower percentages of CD13+ myeloid progeny (compare panels D and E). Only 2 cultures initiated with ALDHbr CD34neg cells had growth significant enough to evaluate lineage development, and these gave rise to CD56+ progeny (C,F). The CD56+ progeny of ALDHneg CD34+ (G-I) and ALDHbr CD34+ cells (data not shown) expressed other antigens consistent with NK cells. (J) Relative output of lymphoid cells was higher in cultures initiated with ALDHneg CD34+ cells when compared with paired cultures initiated with ALDHbr CD34+ cells. (K) Conversely, the relative cell output of myeloid cells was higher in cultures initiated with ALDHbr CD34+ cells when compared with paired cultures initiated with ALDHneg CD34+ cells. Estimations for relative cell output are described in “Materials and methods.” (J-K) Each data point represents an average from duplicate cultures.

Cell development in short-term culture assays. ALDHneg CD34+, ALDHbr CD34+, and ALDHbr CD34neg cells were purified from linneg SSClo UCB, as depicted in Figure 1. Two hundred to 1000 purified cells were cultured on STO fibroblasts in the presence of IL-3, IL-7, and IL-15 (n = 10). Cultures initiated with ALDHneg CD34+ cells contained higher percentages of CD56+ lymphoid progeny than did the ALDHbr CD34+ cells (compare panels A and B; see Table 4). Similarly, the ALDHneg CD34+ cell fraction yielded lower percentages of CD13+ myeloid progeny (compare panels D and E). Only 2 cultures initiated with ALDHbr CD34neg cells had growth significant enough to evaluate lineage development, and these gave rise to CD56+ progeny (C,F). The CD56+ progeny of ALDHneg CD34+ (G-I) and ALDHbr CD34+ cells (data not shown) expressed other antigens consistent with NK cells. (J) Relative output of lymphoid cells was higher in cultures initiated with ALDHneg CD34+ cells when compared with paired cultures initiated with ALDHbr CD34+ cells. (K) Conversely, the relative cell output of myeloid cells was higher in cultures initiated with ALDHbr CD34+ cells when compared with paired cultures initiated with ALDHneg CD34+ cells. Estimations for relative cell output are described in “Materials and methods.” (J-K) Each data point represents an average from duplicate cultures.

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