Cell Therapy for Cystic Fibrosis Lung Disease: Regenerative Basal Cell Amplification
Cell Therapy for Cystic Fibrosis Lung Disease: Regenerative Basal Cell Amplification
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DOI：
10.1002/sctm.18-0098
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发表时间：
2019-03-01
影响因子：
6
通讯作者：
Reynolds, Susan D.
中科院分区：
文献类型：
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作者：
Hayes, Don, Jr.;Kopp, Benjamin T.;Reynolds, Susan D.
人气道上皮细胞由基底细胞再生。因此，基底细胞疗法具有治愈囊性纤维化（CF）肺病的潜力。我们先前报道了移植后人类基底细胞重新填充小鼠气道上皮，我们估计需要6000万个细胞来治疗人类患者。为了进一步发展细胞疗法，我们比较了非CF和CF组织来源的支气管基底细胞的增殖潜力。使用了三种方法：再生细胞频率、爆发大小和细胞分裂频率。其次，我们使用连续传代策略来确定CF基底细胞是否可以扩增至估计的治疗剂量。这些研究评价了组织来源的支气管基底细胞和通过刷支气管气道或鼻呼吸道上皮细胞恢复的基底细胞。最后，我们使用有限稀释法分离非CF和CF基底细胞克隆。增殖试验和气-液-界面分化方法用于确定细胞扩增是否改变克隆分离株的增殖和/或分化潜力。我们证明：（a）非CF和CF基底细胞增殖相似，（B）CF基底细胞可扩增至治疗性细胞剂量，和（c）扩增的非CF和CF基底细胞克隆正常分化。尽管有这些令人鼓舞的发现，我们也发现细胞扩增过程耗尽再生的基底细胞池。基底细胞克隆的分析表明，连续传代选择长寿命的基底细胞，并提高了这些干细胞样细胞的前瞻性分离将提高细胞替代疗法的疗效的可能性。
The human airway epithelium is regenerated by basal cells. Thus, basal cell therapy has the potential to cure cystic fibrosis (CF) lung disease. We previously reported that the human basal cells repopulated the mouse airway epithelium after transplantation, and we estimated that 60 million cells would be needed to treat a human patient. To further develop cell therapy, we compared the proliferation potential of non-CF and CF tissue-derived bronchial basal cells. Three methods were used: regenerative cell frequency, burst size, and cell division frequency. Second, we used a serial passage strategy to determine if CF basal cells could be amplified to the estimated therapeutic dose. These studies evaluated that tissue-derived bronchial basal cells and the basal cells that were recovered by brushing bronchial airways or the nasal respiratory epithelium. Finally, we used the limiting dilution method to isolate non-CF and CF basal cell clones. The proliferation assays and the air-liquid-interface differentiation method were used to determine if cell amplification altered the proliferation and/or differentiation potential of clonal isolates. We demonstrate that: (a) non-CF and CF basal cell proliferation is similar, (b) CF basal cells can be amplified to a therapeutic cell dose, and (c) amplified non-CF and CF basal cell clones differentiate normally. Despite these encouraging findings, we also find that the cell amplification process depletes the regenerative basal cell pool. Analysis of basal cell clones indicates that serial passage selects for long-lived basal cells and raise the possibility that prospective isolation of these stem-like cells will improve the efficacy of cell replacement therapy.