Multiplexed evaluation of mouse wound tissue using oligonucleotide barcoding with single-cell RNA sequencing.
Multiplexed evaluation of mouse wound tissue using oligonucleotide barcoding with single-cell RNA sequencing.
复制标题
使用单细胞RNA测序的寡核苷酸条形码对小鼠伤口组织的多路复用评估。
DOI：
10.1016/j.xpro.2022.101946
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发表时间：
2023-03-17
期刊：
影响因子：
--
通讯作者：
Longaker, Michael T.
中科院分区：
文献类型：
--
作者：
Januszyk, Michael;Griffin, Michelle;Mascharak, Shamik;Talbott, Heather E.;Chen, Kellen;Henn, Dominic;Spielman, Amanda F.;Parker, Jennifer B. L.;Liang, Norah E.;Cotterell, Asha;Guardino, Nicholas;Foster, Deshka S.;Wagh, Dhananjay;Coller, John;Gurtner, Geoffrey C.;Wan, Derrick C.;Longaker, Michael T.
尽管单细胞 RNA 测序在复杂组织研究中的可用性迅速提高，但对于大型研究来说仍然昂贵得令人望而却步。在这里，我们提出了一种使用寡核苷酸条形码来标记和汇集来自愈合伤口的多个样本的协议，这些样本是该应用中最具挑战性的组织类型之一。我们描述了在小鼠中产生皮肤伤口的步骤，然后是组织收获和寡核苷酸条形码。该协议也适用于其他物种，包括大鼠、猪和人类。有关该协议的使用和执行的完整详细信息，请参阅 Stoeckius 等人。 （2018），加利亚诺等人。 (2004) 和 Mascharak 等人。 （2022）。用于标记和合并多个 scRNA-seq 样本的寡核苷酸条形码 使用夹板切除模型进行小鼠伤口愈合 在整个条形码过程中优化组织处理以提高细胞活力 出版商注：进行任何实验方案都需要遵守当地实验室安全和道德机构指南。尽管单细胞 RNA 测序在复杂组织研究中的可用性迅速提高，但对于大型研究来说仍然昂贵得令人望而却步。在这里，我们提出了一种使用寡核苷酸条形码来标记和汇集来自愈合伤口的多个样本的协议，这些样本是该应用中最具挑战性的组织类型之一。我们描述了在小鼠中产生皮肤伤口的步骤，然后是组织收获和寡核苷酸条形码。该协议也适用于其他物种，包括大鼠、猪和人类。
Despite its rapidly increased availability for the study of complex tissue, single-cell RNA sequencing remains prohibitively expensive for large studies. Here, we present a protocol using oligonucleotide barcoding for the tagging and pooling of multiple samples from healing wounds, which are among the most challenging tissue types for this application. We describe steps to generate skin wounds in mice, followed by tissue harvest and oligonucleotide barcoding. This protocol is also applicable to other species including rats, pigs, and humans. For complete details on the use and execution of this protocol, please refer to Stoeckius et al. (2018), Galiano et al. (2004), and Mascharak et al. (2022). Oligonucleotide barcoding for the tagging and pooling of multiple scRNA-seq samples Mouse wound healing using a splinted excisional model Optimized tissue processing for cell viability throughout barcoding process Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Despite its rapidly increased availability for the study of complex tissue, single-cell RNA sequencing remains prohibitively expensive for large studies. Here, we present a protocol using oligonucleotide barcoding for the tagging and pooling of multiple samples from healing wounds, which are among the most challenging tissue types for this application. We describe steps to generate skin wounds in mice, followed by tissue harvest and oligonucleotide barcoding. This protocol is also applicable to other species including rats, pigs, and humans.
影响因子：
23.9
作者：
Mascharak S;Talbott HE;Januszyk M;Griffin M;Chen K;Davitt MF;Demeter J;Henn D;Bonham CA;Foster DS;Mooney N;Cheng R;Jackson PK;Wan DC;Gurtner GC;Longaker MT
通讯作者：
Longaker MT
影响因子：
12.3
作者：
Stoeckius, Marlon;Zheng, Shiwei;Satija, Rahul
通讯作者：
Satija, Rahul
影响因子：
2.9
作者：
Galiano, RD;Michaels, J;Gurtner, GC
通讯作者：
Gurtner, GC