Shibuya A, Nagayoshi K, Nakamura K, Nakauchi H
Division of Hematology, University of Tsukuba, Japan.
Blood. 1995 Jun 15;85(12):3538-46.
We have established a cell culture system without stromal cells that allows the CD34+ hematopoietic progenitor cells (HPC) to differentiate into natural killer (NK) cells. CD34+Lin (CD3, CD16, CD56)- cells were purified using fluorescence-activated cell sorting from normal adult bone marrow (BM) and cultured for 28 days in medium supplemented with interleukin-2 (IL-2) and stem cell factor (SCF). NK (CD3-CD16-CD56+) cells were generated in a dose-dependent manner in response to SCF. NK cells originated from CD34+CD33+Lin- cells, but they were barely detectable in cultures of CD34+CD33-Lin- cells. However, on addition of IL-3, an induced differentiation of NK cells from CD34+CD33-Lin- cells was observed, although at a lower frequency. Supplementing of the cell cultures with SCF alone or both SCF and IL-3 for the first 7 days followed by IL-2 for the next 21 days is essential for production of NK cells from CD34+CD33+Lin- cells and from CD34+CD33-Lin- cells, respectively. These data provide direct evidence that NK cells arise from CD34+HPC and show the minimum lymphokine requirement for their differentiation.
我们建立了一种无基质细胞的细胞培养系统，该系统可使CD34⁺造血祖细胞（HPC）分化为自然杀伤（NK）细胞。使用荧光激活细胞分选技术从正常成人骨髓（BM）中纯化出CD34⁺Lin⁻（CD3、CD16、CD56⁻）细胞，并在添加白细胞介素-2（IL-2）和干细胞因子（SCF）的培养基中培养28天。NK（CD3⁻CD16⁻CD56⁺）细胞以剂量依赖的方式对SCF作出反应而产生。NK细胞起源于CD34⁺CD33⁺Lin⁻细胞，但在CD34⁺CD33⁻Lin⁻细胞培养物中几乎检测不到。然而，添加IL-3后，观察到CD34⁺CD33⁻Lin⁻细胞诱导分化为NK细胞，尽管频率较低。对于分别从CD34⁺CD33⁺Lin⁻细胞和CD34⁺CD33⁻Lin⁻细胞产生NK细胞而言，在最初7天单独用SCF或同时用SCF和IL-3补充细胞培养物，随后在接下来21天用IL-2补充是必不可少的。这些数据提供了直接证据，表明NK细胞起源于CD34⁺HPC，并显示了其分化所需的最低细胞因子要求。
