Multiplexed microfluidic platform for stem-cell derived pancreatic islet β cells.
Multiplexed microfluidic platform for stem-cell derived pancreatic islet β cells.
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DOI：
10.1039/d2lc00468b
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发表时间：
2022-11-08
期刊：
影响因子：
6.1
通讯作者：
Healy, Kevin E.
中科院分区：
文献类型：
--
作者：
Goswami, Ishan;de Klerk, Eleonora;Carnese, Phichitpol;Hebrok, Matthias;Healy, Kevin E.
干细胞来源的β细胞为生物医学发现提供了原始胰岛的替代方案，也为胰岛移植提供了潜在的替代品。获得和维护功能性干细胞来源的β细胞的费用和挑战要求开发更好的高含量和高通量培养系统。微生理系统(MPS)是很有前途的高含量体外平台，但高通量筛选和发现的规模仍然是一个挑战。传统上，液体处理和单元装载的同时多路传输在高通量MPS的设计中是一个挑战。此外，尽管已经开发了用于胰岛β培养/检测的MPS，但在MPS中干细胞来源的β细胞的多天培养的研究一直很有限。我们提出了一种可扩展的、多路复用的胰岛βMPS设备，该设备结合了微流控梯度发生器，以并行处理培养和测试条件下的液体。我们在一周的时间里展示了干细胞来源的富集型β簇(EBCs)的活性和功能，通过簇对葡萄糖的挑战来评估∼2倍的胰岛素释放。为了显示药物测试的可扩展多路传输，我们演示了长期暴露于对数浓度范围的格列本脲后刺激指数的损失。MPS培养的EBCs对代谢产物琥珀酸甲酯和甘油酸的胰岛素分泌反应也证实了糖酵解的瓶颈。因此，我们为EBCS提供了一个平衡高内容和高通量特征的创新文化平台。多路微生理系统作为一种高含量、高通量的设备，用于干细胞来源的β细胞培养和药物筛选。
Stem cell-derived β cells offer an alternative to primary islets for biomedical discoveries as well as a potential surrogate for islet transplantation. The expense and challenge of obtaining and maintaining functional stem cell-derived β cells calls for a need to develop better high-content and high-throughput culture systems. Microphysiological systems (MPS) are promising high-content in vitro platforms, but scaling for high-throughput screening and discoveries remain a challenge. Traditionally, simultaneous multiplexing of liquid handling and cell loading poses a challenge in the design of high-throughput MPS. Furthermore, although MPS for islet β culture/testing have been developed, studies on multi-day culture of stem-cell derived β cells in MPS have been limited. We present a scalable, multiplexed islet β MPS device that incorporates microfluidic gradient generators to parallelize fluid handling for culture and test conditions. We demonstrated the viability and functionality of the stem cell-derived enriched β clusters (eBCs) for a week, as assessed by the ∼2 fold insulin release by the clusters to glucose challenge. To show the scalable multiplexing for drug testing, we demonstrated the loss of stimulation index after long-term exposure to logarithmic concentration range of glybenclamide. The MPS cultured eBCs also confirmed a glycolytic bottleneck as inferred by insulin secretion responses to metabolites methyl succinate and glyceric acid. Thus, we present an innovative culture platform for eBCs with a balance of high-content and high-throughput characteristics. Multiplexed microphysiological system as a high-content, higher throughput device for stem cell-derived β cell culture and drug screening.
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1型糖尿病患者的干细胞衍生的β细胞产生。
影响因子：
16.6
作者：
Millman JR;Xie C;Van Dervort A;Gürtler M;Pagliuca FW;Melton DA
通讯作者：
Melton DA
影响因子：
7.4
作者：
Dishinger, John F.;Reid, Kendra R.;Kennedy, Robert T.
通讯作者：
Kennedy, Robert T.
影响因子：
4.6
作者：
Ionescu-Tirgoviste C;Gagniuc PA;Gubceac E;Mardare L;Popescu I;Dima S;Militaru M
通讯作者：
Militaru M
影响因子：
5.2
作者：
de Klerk E;Hebrok M
通讯作者：
Hebrok M
DOI：
10.1038/s41574-021-00568-0
发表时间：
2022-01
期刊：
Nature reviews. Endocrinology
影响因子：
--
作者：
Benninger RKP;Kravets V
通讯作者：
Kravets V