Primitive long-term culture initiating cells (LTC-ICs) in granulocyte colony-stimulating factor mobilized peripheral blood progenitor cells have similar potential for ex vivo expansion as primitive LTC-ICs in steady state bone marrow.
Primitive long-term culture initiating cells (LTC-ICs) in granulocyte colony-stimulating factor mobilized peripheral blood progenitor cells have similar potential for ex vivo expansion as primitive LTC-ICs in steady state bone marrow.
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DOI：
10.1182/blood.v89.11.3991
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发表时间：
1997-06
期刊：
影响因子：
20.3
通讯作者：
Felipe Prósper;Kirk Vanoverbeke;David F. Stroncek;Catherine M. Verfaillie
中科院分区：
文献类型：
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作者：
Felipe Prósper;Kirk Vanoverbeke;David F. Stroncek;Catherine M. Verfaillie
我们最近发现，正常人外周血中动员的长期培养起始细胞（LTC-IC）中有90%以上表达HLA-DR和CD 38抗原，并且只能维持造血5周。而动员后外周血中10%的LTC-IC为CD 34 + HLA-DR-和CD 34 + CD 38-，可维持造血至少8周。我们现在研究了在粒细胞集落刺激因子（G-CSF）动员的PB祖细胞（PBPC）中CD 34 + HLA-DR+细胞（富含成熟LTC-IC）和CD 34 + HLA-DR-细胞（富含原始LTC-IC）的体外扩增潜力。将细胞与M2- 10 B4细胞接触（接触）或在M2- 10 B4上方的transwell中（非接触）培养2周和5周，其中不含和含白细胞介素-3（IL-3）和巨噬细胞炎性蛋白（MIP-1 α）。评价子代中是否存在集落形成细胞（CFC）和LTC-IC。当CD 34 + HLA-DR+ PB细胞在无细胞因子的接触培养物中培养时，在2周时观察到CFC扩增3倍，但在5周时观察到CFC减少80%。此外，LTC-IC在第2周的恢复仅为17%，在第5周为1%。这证实了我们之前的观察，即虽然CD 34 + HLA-DR+动员的PB细胞可以启动长期培养，但它们相对成熟，不能维持长期造血。与此相反，当CD 34 + HLA-DR动员的PB细胞在接触培养无细胞因子培养，CFC扩增持续到第5周和49%和11%的LTC-IC恢复，分别在第2周和第5周。正如我们已经证明的稳态骨髓（BM）祖细胞，当CD 34 + HLA-DR- PBPC在非接触培养中而不是在接触培养中培养时，LTC-IC的回收率高出3倍，并且当IL-3和MIP-1 α加入非接触培养中时，LTC-IC的回收率进一步提高（第5周时维持在96 +/-2%）。我们的结论是，虽然G-CSF动员了大量的具有有限增殖能力的“成熟”CD 34 + HLA-DR+ LTC-IC，但动员的PB中存在的原始CD 34 + HLA-DR- LTC-IC具有与稳态BM中的LTC-IC相似的特征：（1）它们可以在含有IL-3和MIP-1 α的非接触培养物中维持至少5周;（2）它们受到与基质接触的相同的增殖抑制影响。由于每mL G-CSF动员的PB中原始LTC-IC（第8周LTC-IC）的绝对数与每mL稳态BM中的绝对数相似，因此这些研究进一步证实G-CSF动员的PBPC可能具有与稳态BM相似的长期再增殖能力。
We have recently shown that more than 90% of long-term culture initiating cells (LTC-IC) mobilized in the peripheral blood (PB) of normal individuals express HLA-DR and CD38 antigens and can sustain hematopoiesis for only 5 weeks. However, 10% of LTC-IC in mobilized PB are CD34+ HLA-DR- and CD34+ CD38- and can sustain hematopoiesis for at least 8 weeks. We now examine the ex vivo expansion potential of CD34+ HLA-DR+ cells (rich in mature LTC-IC) and CD34+ HLA-DR- cells (rich in primitive LTC-IC) in granulocyte colony-stimulating factor (G-CSF) mobilized PB progenitor cells (PBPC). Cells were cultured in contact with M2-10B4 cells (contact) or in transwells above M2-10B4 (noncontact) without and with interleukin-3 (IL-3) and macrophage inflammatory protein (MIP-1alpha) for 2 and 5 weeks. Progeny were evaluated for the presence of colony-forming cells (CFC) and LTC-IC. When CD34+ HLA-DR+ PB cells were cultured in contact cultures without cytokines, a threefold expansion of CFC was seen at 2 weeks, but an 80% decrease in CFC was seen at week 5. Further, the recovery of LTC-IC at week 2 was only 17% and 1% at week 5. This confirms our previous observation that although CD34+ HLA-DR+ mobilized PB cells can initiate long-term cultures, they are relatively mature and cannot sustain long-term hematopoiesis. In contrast, when CD34+ HLA-DR- mobilized PB cells were cultured in contact cultures without cytokines, CFC expansion persisted until week 5 and 49% and 11% of LTC-IC were recovered at week 2 and 5, respectively. As we have shown for steady state bone marrow (BM) progenitors, recovery of LTC-IC was threefold higher when CD34+ HLA-DR- PBPC were cultured in noncontact rather than contact cultures, and improved further when IL-3 and MIP-1alpha were added to noncontact cultures (96 +/- 2% maintained at week 5). We conclude that although G-CSF mobilizes a large population of "mature" CD34+ HLA-DR+ LTC-IC with a limited proliferative capacity, primitive CD34+ HLA-DR- LTC-IC present in mobilized PB have similar characteristics as LTC-IC from steady state BM: (1) they can be maintained in noncontact cultures containing IL-3 and MIP-1alpha for at least 5 weeks; (2) they are subject to the same proliferation inhibitory influences of contact with stroma. Since the absolute number of primitive LTC-IC (week 8 LTC-IC) per mL of G-CSF mobilized PB is similar to that per mL of steady state BM, these studies further confirm that G-CSF mobilized PBPC may have similar long-term repopulating abilities as steady state BM.