Hirschhorn R, Ellenbogen A, Martiniuk F
J Cell Physiol. 1985 May;123(2):277-82. doi: 10.1002/jcp.1041230219.
We have substituted deoxyadenosine or adenosine for hypoxanthine in the standard HAT selection system in an attempt to select for ADA-normal (ADA+) cells. ADA- human lymphoid line cells could not utilize deoxyadenosine as an alternative to hypoxanthine as a purine source (DAT) and failed to grow but were only somewhat inhibited in growth when adenosine was substituted for hypoxanthine (AAT). In contrast, ADA+ cells utilized adenosine or deoxyadenosine as efficiently as hypoxanthine as a purine source. Growth in DAT, but not in HAT, of an artificial mixture of one ADA+ human lymphoid cells in 1,000 ADA- cells resulted in enrichment of ADA+ cells to 25-86% of total cells. When we grew a rat ADA- cell line in two variations of the DAT system, we detected at least three electrophoretically different ADA+ patterns, one of which corresponded to normal rat ADA. These could represent "revertants."
我们在标准的HAT选择系统中用脱氧腺苷或腺苷替代次黄嘌呤，试图筛选出腺苷脱氨酶正常（ADA+）的细胞。ADA-人淋巴系细胞不能利用脱氧腺苷替代次黄嘌呤作为嘌呤来源（DAT），无法生长，但当用腺苷替代次黄嘌呤（AAT）时，生长仅受到一定程度的抑制。相比之下，ADA+细胞利用腺苷或脱氧腺苷作为嘌呤来源的效率与利用次黄嘌呤一样高。在1000个ADA-细胞中加入1个ADA+人淋巴系细胞的人工混合物，在DAT中而非HAT中生长，可使ADA+细胞富集至总细胞数的25%-86%。当我们在DAT系统的两种变体中培养大鼠ADA-细胞系时，检测到至少三种电泳图谱不同的ADA+模式，其中一种与正常大鼠ADA相对应。这些可能代表“回复体”。
