Shapiro F, Yao T J, Raptis G, Reich L, Norton L, Moore M A
James Ewing Laboratory of Developmental Hematopoiesis, Memorial Sloan-Kettering Cancer Center, New York, NY 10021.
Blood. 1994 Nov 15;84(10):3567-74.
Multiple cycles of high-dose chemotherapy can be hematologically supported by repeated administration of peripheral blood progenitors obtained after mobilization using cytokine alone or in combination with chemotherapy. We have explored the quality of such cells and their potential to undergo ex vivo expansion. Twenty-five leukapheresis samples from 19 patients who had received extensive prior chemotherapy for stage IV breast cancer were subjected to CD34+ cell selection using immunoaffinity columns of immunomagnetic bead separation. Cells were cultured in suspension in the presence of c-kit ligand, interleukin-3, interleukin-6, erythropoietin, and granulocyte colony-stimulating factor. Ten experiments were performed using weekly exchange of media and cytokines (Delta assay). Median myeloid and erythroid progenitors expanded 15-fold at 7 days (range, 7 to 43), 40-fold at 14 days (range, 18 to 470), 46-fold at 21 days (range, 0 to 118), and 21-fold at 28 days (range, 0 to 61). In a system using gas-permeable bags without exchange of media or cytokine, median progenitors expanded 13-fold at 7 days (range, 7 to 36), 14-fold at 10 days (range, 4 to 61), 14-fold at 12 days (range, 3 to 46), and 10-fold at 14 days (range, 1 to 35). Progenitor expansion less than 10-fold occurred in 8% of experiments at day 7, in 17% at day 10, in 43% at day 12, and in 50% at day 14. When autologous plasma, autologous plasma processed (removal of cryoprecipitate, centrifugation, then filtration), or human serum were substituted for 20% fetal calf serum, the ratio of progenitor expansion at 7 days relative to 20% fetal calf serum for 10% human serum, 20% human serum, and 1% autologous plasma processed was 1.01 (range, 0.62 to 1.33), 0.88 (range, 0.61 to 1.20), and 0.96 (range, 0.55 to 1.64), respectively. These findings support the feasibility of ex vivo expansion in a system free of nonhuman proteins of CD34(+)-derived progenitors obtained from the peripheral blood of patients who have received prior chemotherapy.
多次高剂量化疗周期可通过重复给予单用人细胞因子或联合化疗动员后获得的外周血祖细胞来提供血液学支持。我们研究了此类细胞的质量及其体外扩增的潜力。对19例因IV期乳腺癌接受过广泛前期化疗的患者采集的25份白细胞分离样本，使用免疫磁珠分离免疫亲和柱进行CD34+细胞分选。细胞在含有c-kit配体、白细胞介素-3、白细胞介素-6、促红细胞生成素和粒细胞集落刺激因子的悬浮液中培养。进行了10次实验，每周更换培养基和细胞因子（Delta检测）。髓系和红系祖细胞的中位数在7天时扩增15倍（范围7至43），14天时扩增40倍（范围18至470），21天时扩增46倍（范围0至118），28天时扩增21倍（范围0至61）。在使用透气袋且不更换培养基或细胞因子的系统中，祖细胞中位数在7天时扩增13倍（范围7至36），10天时扩增14倍（范围4至61），12天时扩增14倍（范围3至46），14天时扩增10倍（范围1至35）。祖细胞扩增倍数小于10倍的情况在7天时的实验中占8%，10天时占17%，12天时占43%，14天时占50%。当用自体血浆、处理过的自体血浆（去除冷沉淀、离心，然后过滤）或人血清替代20%胎牛血清时，相对于20%胎牛血清，10%人血清、20%人血清和1%处理过的自体血浆在7天时的祖细胞扩增比例分别为1.01（范围0.62至1.33）、0.88（范围0.61至1.20）和0.96（范围0.55至1.64）。这些发现支持了在无非人源蛋白质的系统中对接受过前期化疗患者外周血中获得的CD34(+)衍生祖细胞进行体外扩增的可行性。
