Kawano Yutaka, Kobune Masayoshi, Chiba Hiroki, Nakamura Kiminori, Takimoto Rishu, Takada Kohichi, Ito Yoshinori, Kato Junji, Hamada Hirofumi, Niitsu Yoshiro
Fourth Dept. of Internal Medicine, Sapporo Medical University School of Medicine, Sapporo, Japan.
Exp Hematol. 2006 Feb;34(2):150-8. doi: 10.1016/j.exphem.2005.10.007.
The pentaspan molecule CD133 has been shown to be a marker of more primitive hematopoietic progenitors in mobilized peripheral blood (PB). Our objective was to assess the efficacy of PB CD133(+) cells in our coculture system using human telomerized stromal (HTS) cells.
Five thousand PB CD133(+) cells or conventional cord blood (CB) CD34(+) cells were expanded with or without HTS cells in the presence or absence of stem cell factor, thrombopoietin, and Flk-2/Flt-3 ligand.
The coculture was significantly superior in expanding PB clonogenic cells as compared with the stroma-free culture (CFU-C, 2 +/- 0 vs 111 +/- 15-fold of initial cell number, p < 0.01), and the fold increase of PB clonogenic cells was comparable to that for CB cells after two weeks of coculture (BFU-E, 54 +/- 3 vs 56 +/- 4-fold; CFU-GM, 156 +/- 26 vs 83 +/- 9-fold; CFU-Mix, 30 +/- 11 vs 80 +/- 36-fold). However, proliferation of CFU-Mk from PB on coculture with HTS cells was modest as compared with stroma-free culture. Concomitantly, multiple hematopoietic cells transmigrated below the stromal layer and formed cobblestone areas (CAs). The production of hematopoietic progenitor cells from CAs after coculture with PB was significantly lower than that seen in cells cocultured with CB for four weeks (CFU-Mix, 0 +/- 0 vs 9 +/- 5-fold on day 28, p < 0.01), although a similar number of CAs derived from PB and CB were observed.
PB CD133(+) cells proliferated efficiently above the stromal layer, while the characteristics of PB CD133(+) cells underneath the human stromal layer were likely to be maintained, even after long-term hematopoietic-stromal interaction.
五跨膜分子CD133已被证明是动员外周血（PB）中更原始造血祖细胞的标志物。我们的目的是评估在我们使用人端粒化基质（HTS）细胞的共培养系统中PB CD133(+)细胞的效能。
5000个PB CD133(+)细胞或传统脐带血（CB）CD34(+)细胞在有或无HTS细胞的情况下，于有或无干细胞因子、血小板生成素和Flk-2/Flt-3配体存在时进行扩增。
与无基质培养相比，共培养在扩增PB集落形成细胞方面显著更优（CFU-C，初始细胞数的2±0倍与111±15倍，p<0.01），并且共培养两周后PB集落形成细胞的增加倍数与CB细胞相当（BFU-E，54±3倍与56±4倍；CFU-GM，156±26倍与83±9倍；CFU-Mix，30±11倍与80±36倍）。然而，与无基质培养相比，PB与HTS细胞共培养时CFU-Mk的增殖较为适度。同时，多种造血细胞迁移至基质层下方并形成鹅卵石区（CA）。PB与CA共培养后造血祖细胞的产生显著低于与CB共培养四周时所见（CFU-Mix，第28天为0±0倍与9±5倍，p<0.01），尽管观察到来自PB和CB的CA数量相似。
PB CD133(+)细胞在基质层上方高效增殖，而即使经过长期造血-基质相互作用，人基质层下方PB CD133(+)细胞的特性可能仍得以维持。
