Optimized isolation and expansion of human airway epithelial basal cells from endobronchial biopsy samples.
Optimized isolation and expansion of human airway epithelial basal cells from endobronchial biopsy samples.
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DOI：
10.1002/term.2466
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发表时间：
2018-01
影响因子：
3.3
通讯作者：
Janes SM
中科院分区：
文献类型：
--
作者：
Gowers KHC;Hynds RE;Thakrar RM;Carroll B;Birchall MA;Janes SM
自体气道上皮细胞已被用于临床组织工程气道移植手术，以期帮助粘膜再生和恢复粘膜纤毛自动扶梯功能。然而，由于这些干预措施的紧迫性，可用于上皮细胞扩张的时间有限，而且在患者中观察到了缓慢的上皮再生。人的呼吸道上皮细胞可以从支气管镜检查过程中的小活检或刷子中扩增，但从患者身上获取组织的最佳模式尚未被研究。在这里，我们比较了支气管镜下刷检和支气管镜活检标本的细胞数和启动基础上皮干细胞培养的能力。我们发现，在含有Rho相关蛋白激酶抑制剂Y-27632的培养液中，样品与3T3-J2饲养层细胞直接共培养，在关键的早期培养阶段，与传统技术相比，选择性地扩增了更多数量的基底上皮干细胞。此外，我们确定了从细胞悬液中启动细胞培养的好处，无论是使用刷检样本还是通过活组织切片的酶消化，都优于外植体培养。原代上皮细胞培养是从在细胞培养开始之前已经冷冻保存的支气管腔内活检样本开始的，这表明冷冻保存可以消除采集活检样本的临床设施和培养上皮细胞的专业实验室之间距离很近的要求。总体而言，我们的结果建议在未来的呼吸道细胞治疗或生物工程气道移植程序中加快上皮细胞准备的方法。
Autologous airway epithelial cells have been used in clinical tissue‐engineered airway transplantation procedures with a view to assisting mucosal regeneration and restoring mucociliary escalator function. However, limited time is available for epithelial cell expansion due to the urgent nature of these interventions and slow epithelial regeneration has been observed in patients. Human airway epithelial cells can be expanded from small biopsies or brushings taken during bronchoscopy procedures, but the optimal mode of tissue acquisition from patients has not been investigated. Here, we compared endobronchial brushing and endobronchial biopsy samples in terms of their cell number and their ability to initiate basal epithelial stem cell cultures. We found that direct co‐culture of samples with 3T3‐J2 feeder cells in culture medium containing a Rho‐associated protein kinase inhibitor, Y‐27632, led to the selective expansion of greater numbers of basal epithelial stem cells during the critical early stages of culture than traditional techniques. Additionally, we established the benefit of initiating cell cultures from cell suspensions, either using brushing samples or through enzymatic digestion of biopsies, over explant culture. Primary epithelial cell cultures were initiated from endobronchial biopsy samples that had been cryopreserved before the initiation of cell cultures, suggesting that cryopreservation could eliminate the requirement for close proximity between the clinical facility in which biopsy samples are taken and the specialist laboratory in which epithelial cells are cultured. Overall, our results suggest ways to expedite epithelial cell preparation in future airway cell therapy or bioengineered airway transplantation procedures.
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影响因子：
2.1
作者：
DEJONG, PM;VANSTERKENBURG, MAJA;PONEC, M
通讯作者：
PONEC, M
影响因子：
4.6
作者：
Walsh, Alex J.;Cook, Rebecca S.;Skala, Melissa C.
通讯作者：
Skala, Melissa C.
DOI：
10.1165/rcmb.2015-0274ma
发表时间：
2016-09-01
影响因子：
6.4
作者：
Reynolds, Susan D.;Rios, Cydney;Seibold, Max A.
通讯作者：
Seibold, Max A.
影响因子：
6
作者：
Palechor-Ceron, Nancy;Suprynowicz, Frank A.;Liu, Xuefeng
通讯作者：
Liu, Xuefeng
影响因子：
8.8
作者：
Danahay, Henry;Pessotti, Angelica D.;Jaffe, Aron B.
通讯作者：
Jaffe, Aron B.