Reliable multiplex generation of pooled induced pluripotent stem cells.
Reliable multiplex generation of pooled induced pluripotent stem cells.
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DOI：
10.1016/j.crmeth.2023.100570
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发表时间：
2023-09-25
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影响因子：
--
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中科院分区：
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将体细胞重编程为多能干细胞（iPSC）使得能够在体外研究系统。为了提高重编程的通量，我们提出了从合并细胞（iPPC）诱导多能性-一种有效的，可扩展的和可靠的重编程程序。使用我们采用单核苷酸多态性（SNP）的合并测序的去卷积算法，我们准确地估计了合并的iPSC的个体供体比例。使用iPPC，我们同时将一百多个供体淋巴母细胞系（LCL）重编程为iPSC，并在多个实验中发现个体供体重编程能力的强相关性。个体供体的重编程能力在同一天的重复和多次实验运行中保持一致，并且某些免疫球蛋白前体基因的表达可能影响重编程能力。合并的iPSC也能够分化成脑类器官。我们的程序使得能够使用供体iPSC的汇集文库进行下游研究和体外表型的调查的多重框架。多供体LCL可以作为一个池重编程为iPSC个体比例可以很容易地从低覆盖测序中确定个体比例在不同的实验重复中是一致的这种方法有助于基于iPSC的实验模型的多路复用将成熟细胞重编程为诱导多能干细胞（iPSC）的能力是一项重大发现，它使大量的下游实验应用成为可能。然而，重编程效率仍然很低，并且许多不同供体iPSC群体的实验建模是繁琐的。在这里，我们证明了重编程可以作为许多不同供体细胞的混合池进行，并且我们可以通过低通DNA测序来解卷积供体身份及其在池内的各自比例。Smullen等人报道iPPC，一种从多个供体LCL作为池产生iPSC的方法。混合iPSC的个体比例可以通过低覆盖率的下一代测序轻松获得。在不同的实验重复中，比例是一致的。此外，iPSC可以分化成脑类器官。
Reprogramming somatic cells into pluripotent stem cells (iPSCs) enables the study of systems in vitro. To increase the throughput of reprogramming, we present induction of pluripotency from pooled cells (iPPC)—an efficient, scalable, and reliable reprogramming procedure. Using our deconvolution algorithm that employs pooled sequencing of single-nucleotide polymorphisms (SNPs), we accurately estimated individual donor proportions of the pooled iPSCs. With iPPC, we concurrently reprogrammed over one hundred donor lymphoblastoid cell lines (LCLs) into iPSCs and found strong correlations of individual donors’ reprogramming ability across multiple experiments. Individual donors’ reprogramming ability remains consistent across both same-day replicates and multiple experimental runs, and the expression of certain immunoglobulin precursor genes may impact reprogramming ability. The pooled iPSCs were also able to differentiate into cerebral organoids. Our procedure enables a multiplex framework of using pooled libraries of donor iPSCs for downstream research and investigation of in vitro phenotypes. Multidonor LCLs can be reprogrammed into iPSCs as a pool The individual proportions can be easily determined from low-coverage sequencing The individual proportions are consistent across different experimental replicates This method contributes to the multiplexing of experimental iPSC-based models The ability to reprogram mature cells into induced pluripotent stem cells (iPSCs) was a major discovery that enabled a plethora of downstream experimental applications. However, the reprogramming efficiency remains low, and experimental modeling of a population of many different donor iPSCs is tedious. Here, we demonstrate that reprogramming can be performed as a mixed pool of many different donor cells, and we can deconvolute donor identities and their respective proportions within the pool via low-pass DNA sequencing. Smullen et al. report iPPC, a method for generating iPSCs from multiple donor LCLs as a pool. The individual proportions of the mixed iPSCs can be easily accessed via low-coverage next-generation sequencing. The proportions are consistent across different experimental replicates. Also, the iPSCs can be differentiated into cerebral organoids.
改进的方法，使用荧光激活的细胞分选来重编程人皮肤成纤维细胞。
影响因子：
3.7
作者：
Kahler DJ;Ahmad FS;Ritz A;Hua H;Moroziewicz DN;Sproul AA;Dusenberry CR;Shang L;Paull D;Zimmer M;Weiss KA;Egli D;Noggle SA
通讯作者：
Noggle SA
影响因子：
12.3
作者：
Chan Y;Chan YK;Goodman DB;Guo X;Chavez A;Lim ET;Church GM
通讯作者：
Church GM