Haploid embryonic stem cells can be enriched and maintained by simple filtration | |
Qu, Chao1; Yan, Meng1; Yang, Suming1; Wang, Lingbo1; Yin, Qi1; Li, Jinsong1![]() | |
刊名 | JOURNAL OF BIOLOGICAL CHEMISTRY
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2018 | |
卷号 | 293期号:14页码:5230-5235 |
关键词 | Genetically-modified Mice Efficient Generation Cyto-toxicity Mouse Genome Derivation Hoechst-33342 Technology Systems Half |
ISSN号 | 0021-9258 |
DOI | 10.1074/jbc.RA118.002029 |
文献子类 | Article |
英文摘要 | Mammalian haploid embryonic stem cells (haESCs) serve as a powerful tool for genetic analyses at both the cellular and organismal levels. However, spontaneous diploidization of haESCs limits their use in these analyses. Addition of small molecules to the culture medium to control the cell cycle can slow down diploidization, but cell-sorting methods such as FACS are still required to enrich haploid cells for long-term maintenance in vitro. Here, acting on our observation that haploid and diploidized cells differ in diameter, we developed a simplified filtration method to enrich haploid cells from cultured haESCs. We found that regular cell filtration with this system reliably maintained the haploidy of mouse haESCs for over 30 passages. Importantly, CRISPR/Cas9-mediated knockout and knockin were successfully achieved in the filtered cells, leading to stable haploid cell lines carrying the desired gene modifications. Of note, by injecting haESCs into metaphase II oocytes, we efficiently obtained live mice with the expected genetic traits, indicating that regular filtration maintained the functional integrity of haESCs. Moreover, this filtration system was also feasible for derivation of mouse haESCs from parthenogenetic haploid blastocysts and for human haESC maintenance. In conclusion, we have identified a reliable, efficient, and easy-to-handle technique for countering diploidization of haploid cells, a major obstacle in haESC applications. |
电子版国际标准刊号 | 1083-351X |
WOS研究方向 | Biochemistry & Molecular Biology |
语种 | 英语 |
WOS记录号 | WOS:000429409300027 |
内容类型 | 期刊论文 |
版本 | 出版稿 |
源URL | [http://202.127.25.143/handle/331003/3415] ![]() |
专题 | 生化所2018年发文 上海生化细胞研究所_上海生科院生化细胞研究所 |
通讯作者 | Li, Jinsong |
作者单位 | 1.Chinese Acad Sci, Shanghai Key Lab Mol Androl, State Key Lab Cell Biol, Inst Biochem & Cell Biol,Univ Chinese Acad Sci,Ct, Shanghai 200031, Peoples R China; 2.Tsinghua Univ, Sch Life Sci, Tsinghua Peking Ctr Life Sci, State Key Lab Membrane Biol, Beijing 100084, Peoples R China |
推荐引用方式 GB/T 7714 | Qu, Chao,Yan, Meng,Yang, Suming,et al. Haploid embryonic stem cells can be enriched and maintained by simple filtration[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2018,293(14):5230-5235. |
APA | Qu, Chao.,Yan, Meng.,Yang, Suming.,Wang, Lingbo.,Yin, Qi.,...&Chen, Yeguang.(2018).Haploid embryonic stem cells can be enriched and maintained by simple filtration.JOURNAL OF BIOLOGICAL CHEMISTRY,293(14),5230-5235. |
MLA | Qu, Chao,et al."Haploid embryonic stem cells can be enriched and maintained by simple filtration".JOURNAL OF BIOLOGICAL CHEMISTRY 293.14(2018):5230-5235. |
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