ArticleStem cells translational medicine2015
A facile method to establish human induced pluripotent stem cells from adult blood cells under feeder-free and xeno-free culture conditions: a clinically compliant approach.
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Article in Stem cells translational medicine, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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40 citing papers in PubMed, 92 citations in OpenAlex.
- Efficient and robust generation of functional hematopoietic cells from human pluripotent stem cells in albumin-free conditions.Stem cells translational medicine · 2026Article
- Progesterone receptor is constitutively expressed in induced Pluripotent Stem Cells (iPSCs).Stem cell reviews and reports · 2024Article
- iPSC-derived hindbrain organoids to evaluate escitalopram oxalate treatment responses targeting neuropsychiatric symptoms in Alzheimer's disease.Molecular psychiatry · 2024Article
- Polygenic risk for schizophrenia converges on alternative polyadenylation as molecular mechanism underlying synaptic impairment.bioRxiv : the preprint server for biology · 2024Article
- CRISPR-Cas12a for Highly Efficient and Marker-Free Targeted Integration in Human Pluripotent Stem Cells.International journal of molecular sciences · 2024Article
- Avian iPSC Derivation to Recover Threatened Wild Species: A Comprehensive Review in Light of Well-Established Protocols.Animals : an open access journal from MDPI · 2024Review
- Generation and Characterization of a Human-Derived and Induced Pluripotent Stem Cell (iPSC) Line from an Alzheimer's Disease Patient with Neuropsychiatric Symptoms.Biomedicines · 2023Article
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- BdLT-Seq as a barcode decay-based method to unravel lineage-linked transcriptome plasticity.Nature communications · 2023Article
- Defective hematopoietic differentiation of immune aplastic anemia patient-derived iPSCs.Cell death & disease · 2022Article
- Identification of Drug Transporter Genomic Variants and Inhibitors That Protect Against Doxorubicin-Induced Cardiotoxicity.Circulation · 2022Article
- RARG variant predictive of doxorubicin-induced cardiotoxicity identifies a cardioprotective therapy.Cell stem cell · 2021Article
- An Overview on Promising Somatic Cell Sources Utilized for the Efficient Generation of Induced Pluripotent Stem Cells.Stem cell reviews and reports · 2021Review
- Generation of Primordial Germ Cell-like Cells from iPSCs Derived from Turner Syndrome Patients.Cells · 2021Article
- Protocol for generating human induced neural progenitor cells from immobilized adult peripheral blood.STAR protocols · 2021Article
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- Cystatin B is essential for proliferation and interneuron migration in individuals with EPM1 epilepsy.EMBO molecular medicine · 2020Article
- Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture.Stem cell reports · 2020Article
- iPSCs from people with MS can differentiate into oligodendrocytes in a homeostatic but not an inflammatory milieu.PloS one · 2020Article
- Episomal Induced Pluripotent Stem Cells: Functional and Potential Therapeutic Applications.Cell transplantation · 2019Review
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Authors and funding
10 authors at 4 institutions in 2 countries.
Bin-Kuan ChouDivision of Hematology, Department of Medicine, and Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences, and Department of Stem Cells and Regenerative Medicine, Peking Union Medical College, Tianjin, People's Republic of China; Department of Transfusion, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, USA; Key Laboratory of Pediatric Hematology/Oncology of Ministry of Health, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Haihui GuDivision of Hematology, Department of Medicine, and Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences, and Department of Stem Cells and Regenerative Medicine, Peking Union Medical College, Tianjin, People's Republic of China; Department of Transfusion, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, USA; Key Laboratory of Pediatric Hematology/Oncology of Ministry of Health, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Yongxing GaoDivision of Hematology, Department of Medicine, and Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences, and Department of Stem Cells and Regenerative Medicine, Peking Union Medical College, Tianjin, People's Republic of China; Department of Transfusion, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, USA; Key Laboratory of Pediatric Hematology/Oncology of Ministry of Health, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Sarah N DoweyDivision of Hematology, Department of Medicine, and Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences, and Department of Stem Cells and Regenerative Medicine, Peking Union Medical College, Tianjin, People's Republic of China; Department of Transfusion, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, USA; Key Laboratory of Pediatric Hematology/Oncology of Ministry of Health, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Ying WangDivision of Hematology, Department of Medicine, and Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences, and Department of Stem Cells and Regenerative Medicine, Peking Union Medical College, Tianjin, People's Republic of China; Department of Transfusion, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, USA; Key Laboratory of Pediatric Hematology/Oncology of Ministry of Health, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Jun ShiDivision of Hematology, Department of Medicine, and Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences, and Department of Stem Cells and Regenerative Medicine, Peking Union Medical College, Tianjin, People's Republic of China; Department of Transfusion, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, USA; Key Laboratory of Pediatric Hematology/Oncology of Ministry of Health, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Yanxin LiDivision of Hematology, Department of Medicine, and Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences, and Department of Stem Cells and Regenerative Medicine, Peking Union Medical College, Tianjin, People's Republic of China; Department of Transfusion, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, USA; Key Laboratory of Pediatric Hematology/Oncology of Ministry of Health, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Zhaohui YeDivision of Hematology, Department of Medicine, and Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences, and Department of Stem Cells and Regenerative Medicine, Peking Union Medical College, Tianjin, People's Republic of China; Department of Transfusion, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, USA; Key Laboratory of Pediatric Hematology/Oncology of Ministry of Health, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Tao ChengDivision of Hematology, Department of Medicine, and Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences, and Department of Stem Cells and Regenerative Medicine, Peking Union Medical College, Tianjin, People's Republic of China; Department of Transfusion, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, USA; Key Laboratory of Pediatric Hematology/Oncology of Ministry of Health, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Linzhao ChengDivision of Hematology, Department of Medicine, and Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences, and Department of Stem Cells and Regenerative Medicine, Peking Union Medical College, Tianjin, People's Republic of China; Department of Transfusion, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, USA; Key Laboratory of Pediatric Hematology/Oncology of Ministry of Health, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China lcheng2@jhmi.edu.
Johns Hopkins University · USInstitute of Hematology & Blood Diseases Hospital · CNJohns Hopkins Medicine · USShanghai Children's Medical Center · CN
Funding
Functional Genomics of Platelet Aggregation Using iPS and Derived MegakaryocytesU01HL107446 · NHLBI · JOHNS HOPKINS UNIVERSITY · PI BECKER, LEWIS C, CHENG, LINZHAO · 2011 to 2015
$8.8MTRAINING PROGRAM IN HEMATOLOGYT32HL007525 · NHLBI · JOHNS HOPKINS UNIVERSITY · PI ROBERT A BRODSKY · 1985 to 2026
$6.2MHuman embryonic stem (ES) and embroyonic germ (EG) cells to blood and immune cellR01HL073781 · NHLBI · JOHNS HOPKINS UNIVERSITY · PI CHENG, LINZHAO · 2003 to 2013
$4.2MNHLBI NIH HHS 2R01 HL-073781NHLBI NIH HHS R01 HL073781NHLBI NIH HHS T32 HL007525NHLBI NIH HHS U01 HL107446
Abstract
Reprogramming human adult blood mononuclear cells (MNCs) cells by transient plasmid expression is becoming increasingly popular as an attractive method for generating induced pluripotent stem (iPS) cells without the genomic alteration caused by genome-inserting vectors. However, its efficiency is relatively low with adult MNCs compared with cord blood MNCs and other fetal cells and is highly variable among different adult individuals. We report highly efficient iPS cell derivation under clinically compliant conditions via three major improvements. First, we revised a combination of three EBNA1/OriP episomal vectors expressing five transgenes, which increased reprogramming efficiency by ≥10-50-fold from our previous vectors. Second, human recombinant vitronectin proteins were used as cell culture substrates, alleviating the need for feeder cells or animal-sourced proteins. Finally, we eliminated the previously critical step of manually picking individual iPS cell clones by pooling newly emerged iPS cell colonies. Pooled cultures were then purified based on the presence of the TRA-1-60 pluripotency surface antigen, resulting in the ability to rapidly expand iPS cells for subsequent applications. These new improvements permit a consistent and reliable method to generate human iPS cells with minimal clonal variations from blood MNCs, including previously difficult samples such as those from patients with paroxysmal nocturnal hemoglobinuria. In addition, this method of efficiently generating iPS cells under feeder-free and xeno-free conditions allows for the establishment of clinically compliant iPS cell lines for future therapeutic applications.
Indexed as
Cell Culture TechniquesAdultBlood CellsCell DifferentiationCellular ReprogrammingCulture Media, Serum-FreeFeeder CellsFlow CytometryGenetic VectorsHumansInduced Pluripotent Stem CellsLeukocytes, MononuclearVitronectinCulture Media, Serum-FreeVitronectinEpisomal vectorsFeeder-freeHuman blood mononuclear cellsHuman iPS cellsPlasmid expressionReprogrammingXeno-free cell culture
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