{"doi":"10.21769/bioprotoc.3788","title":"Derivation of Induced Pluripotent Stem Cells from Human Fibroblasts Using a Non-integrative System in Feeder-free Conditions","abstract":null,"journal":"BIO-PROTOCOL","year":2020,"id":617435,"datarank":0.36104048104637715,"base_score":2.0794415416798357,"endowment":2.0794415416798357,"self_citation_contribution":0.31191623125197543,"citation_network_contribution":0.049124249794401734,"self_endowment_contribution":0.31191623125197543,"citer_contribution":0.049124249794401734,"corpus_percentile":null,"corpus_rank":null,"citation_count":7,"citer_count":5,"citers_with_citation_signal":4,"citers_with_endowment":4,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1592321,"name":"Sarahi Molina","orcid":null,"position":1,"is_corresponding":false},{"id":1592323,"name":"Adriana Beltran","orcid":null,"position":2,"is_corresponding":false},{"id":1592320,"name":"Alvaro Beltran","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Derivation of Induced Pluripotent Stem Cells from Human Fibroblasts Using a Non-integrative System in Feeder-free Conditions","abstract":"Induced pluripotent stem cells (iPSCs) are genetically reprogrammed somatic cells that exhibit features identical to those of embryonic stem cells (ESCs). Multiple approaches are available to derive iPSCs, among which the Sendai virus is the most effective at reprogramming different cell types. Here we describe a rapid, efficient, safe, and reliable approach to reprogram human fibroblasts into iPSCs that are compatible with future iPSCs uses such as genome editing and differentiation to a transplantable cell type., [摘要] 诱导多能干细胞（iPSCs）是一种经过基因重组的体细胞，具有与胚胎干细胞（ESCs）相同的特性。有多种方法可以获得iPSCs，其中仙台病毒是最有效的重编程不同的细胞类型。在这里，我们描述了一种快速、高效、安全、可靠的方法，将人类成纤维细胞重新编程为与将来iPSCs相兼容的iPSCs，如基因组编辑和分化为可移植细胞类型。 [背景] 诱导多能干细胞（iPSCs）是经过基因重组的成体细胞，其形态和功能特性与胚胎干细胞（esc）非常相似（Takahashi和Yamanaka，2006；Yu等人，2007）。它们不仅为疾病建模提供了一个很好的机会，而且为涉及组织退化的病理学的治疗策略的发展提供了一个很好的机会。此外，iPSCs的承诺依赖于一个安全的可补充的细胞源，来源于化学定义的培养基中，并且没有随机的DNA整合。将体细胞重编程为iPSCs需要强制表达支持多潜能状态的转录因子，包括OCT4、SOX2、KLF4、c-MYC、NANOG和LIN-28（Takahashi和Yamanaka，2006；Takahashi等人，2007；Yu等人，2007）。有多种方法可以将转录因子传递到细胞中，包括那些需要整合到宿主染色体中的方法（Takahashi and Yamanaka，2006；Kane等人，2010）。外源性DNA整合会对移植后细胞的质量和安全性产生不可预测的影响。其他方法包括以DNA为基础的载体（Yu等人，2011年；Weltner等人，2012年），因此减少了整合的可能性，最后是那些不整合到宿主基因组中的，被称为无转基因的载体。无转基因方法包括mRNA（Warren and Wang，2013）、重组蛋白（Zhou et al.，2009）和仙台病毒（Fusaki et al.，2009）。以mRNA或重组蛋白的形式传递多能性转录因子是一种低效且昂贵的方法。相比之下，仙台病毒是一种高效的RNA病毒，能有效地对不同类型的体细胞进行重组。仙台病毒（Sendai）是一种胞质RNA复制不全病毒（SeV），它安全有效地将重编程因子传递到体细胞中，不会整合到基因组或改变细胞的遗传信息（Li等人，2000；Fusaki等人，2009）。此外，病毒在经过几次传代后被清除出细胞，保证了载体和转基因的零足迹。仙台重编程系统的市面上可作为CytoTune iPS 2.0进行研究，也可作为CTS CytoTune iPS 2.1用于临床应用。这两种系统都包含编码四种山中因子（OCT4、SOX2、KLF4和c-MYC）的载体，这些因子被优化用于从人类体细胞生成iPSCs，这使得仙台病毒成为产生无转基因iPSCs的最快速、最有效和最具成本效益的方法。人类iPSCs的可靠和安全的衍生依赖于所定义和合格的试剂的使用，这些试剂允许顺利过渡到下游技术，并且符合大规模细胞生产的GMP（良好制造规范）质量标准。在这里，我们描述了一种人类成纤维细胞的重编程方法，它使用非整合系统，化学定义的培养基，在无饲养条件下。这种方法能够快速、高效、安全、可靠地衍生出与未来应用兼容的iPSCs，包括基因组编辑和向可移植细胞类型分化。","is_dataset_classified":null,"base_score":2.0794415416798357,"endowment":2.0794415416798357,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"33659443","pmcid":"PMC7842795","openalex_id":"https://openalex.org/W3094155397","authors":[],"funders":[],"total_grants":0,"fwci":0.2982,"citation_percentile":0.5322174,"influential_citations":0,"citation_trend":[{"year":2022,"count":2},{"year":2023,"count":2},{"year":2025,"count":3}],"oa_status":"hybrid","license":"cc-by","oa_locations":[{"url":"https://bio-protocol.org/pdf/Bio-protocol3788.pdf","host_type":"journal"},{"url":"https://bio-protocol.org/pdf/Bio-protocol3788.pdf","host_type":"publisher"},{"url":"https://bio-protocol.org/e3788","host_type":"publisher"},{"url":"https://doi.org/10.21769/bioprotoc.3788","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/33659443","host_type":"repository"},{"url":"https://doaj.org/article/ef9bc81bb86c49ac9f255b968e4f0b00","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/7842795","host_type":"repository"}],"fields_of_study":["Pluripotent Stem Cells Research","CRISPR and Genetic Engineering","Biomedical Ethics and Regulation"],"mesh_terms":[],"keywords":["Induced pluripotent stem cell","Reprogramming","Sendai virus","Embryonic stem cell","Somatic cell","Biology","Cell biology","Stem cell","Cell type","Computational biology","Cell","Genetics","Gene","Fibroblasts","Induced Pluripotent Stem Cells","Transgene-free","Feeder-free"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-03T01:42:58.338709Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}