An Efficient Nonviral Method to Generate Integration‐Free Human‐Induced Pluripotent Stem Cells from Cord Blood and Peripheral Blood Cells

  title={An Efficient Nonviral Method to Generate Integration‐Free Human‐Induced Pluripotent Stem Cells from Cord Blood and Peripheral Blood Cells},
  author={Keisuke Okita and Tatsuya Yamakawa and Yasuko Matsumura and Yoshiko Sato and Naoki Amano and Akira Watanabe and Naoki Goshima and Shinya Yamanaka},
  journal={STEM CELLS},
The generation of induced pluripotent stem cells (iPSCs) provides the opportunity to use patient‐specific somatic cells, which are a valuable source for disease modeling and drug discovery. To promote research involving these cells, it is important to make iPSCs from easily accessible and less invasive tissues, like blood. We have recently reported the efficient generation of human iPSCs from adult fibroblasts using a combination of plasmids encoding OCT3/4, SOX2, KLF4, L‐MYC, LIN28, and shRNA… 

Optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood

Improved the generation efficiency of integration-free iPSCs from human peripheral blood mononuclear cells by optimizing the method of isolating mononnuclear cells from peripheral blood, by modifying the integration of culture medium, and by adjusting the duration of culture time and the combination of different episomal vectors.

Advanced Feeder‐Free Generation of Induced Pluripotent Stem Cells Directly From Blood Cells

Efficient generation of iPSCs directly from PBMCs in feeder‐free conditions in approximately 2 weeks are reported, and the produced iPSC clones are pluripotent and transgene‐free, which make this novel method a powerful tool for large‐scale reprogramming ofPBMCs and foriPSC biobanking.

Efficient Generation of Integration-Free iPS Cells from Human Adult Peripheral Blood Using BCL-XL Together with Yamanaka Factors

It is shown that Yamanaka factors (OCT4, SOX2, MYC, and KLF4)-expressing EV can also reprogram adult peripheral blood mononuclear cells (PBMNCs) into pluripotency, yet at a very low efficiency.

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

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, and allows for the establishment of clinically compliant iPS cell lines for future therapeutic applications.

Generation of Induced Pluripotent Stem Cells from Human Peripheral T Cells Using Sendai Virus in Feeder-free Conditions

This protocol demonstrates the generation of iPSCs from human peripheral T cells in feeder-free conditions using a combination of activated T cell culture and Sendai virus to reduce the risk of exposing IPSCs to undefined pathogens.

Reprogramming peripheral blood mononuclear cells using an efficient feeder-free, non-integration method to generate iPS cells and the effect of immunophenotype and epigenetic state on HSPC fate

This project aims to utilize peripheral blood mononuclear cells (MNC), one of the most easily accessible tissues to generate iPS cells using an non-viral, feeder cell free methodology, with the ultimate goal of moving this methodology towards clinical use.

Umbilical Cord Tissue as a Source of Young Cells for the Derivation of Induced Pluripotent Stem Cells Using Non-Integrating Episomal Vectors and Feeder-Free Conditions

It is demonstrated that CT-MSCs can be easily isolated without expensive enzymatic treatment or columns and reprogramed well using episomal vectors, which allow for the removal of the reprogramming factors after a few passages.

Practical Integration‐Free Episomal Methods for Generating Human Induced Pluripotent Stem Cells

This work describes commonly practiced methods for non‐integrating induced pluripotent stem cell generation using nucleofection of episomal reprogramming plasmids, adapted from recent studies that demonstrate increased hiPS cell reprograming efficacy with the application of three powerful episomal hiPS Cell Reprogramming factor vectors.

Derivation, Expansion, and Motor Neuron Differentiation of Human-Induced Pluripotent Stem Cells with Non-Integrating Episomal Vectors and a Defined Xenogeneic-free Culture System

This induction method will promote the derivation of patient-specific integration-free and xeno-free iPSCs and improve the strategy for motor neuron derivation and provides a useful tool for human disease models, drug screen, and clinical applications.

Improvement of transfection with reprogramming factors in urinederived cells

An experimental protocol for obtaining and generating iPS-like cells from urine samples for further cell therapy research on different human diseases is provided and it is concluded that Lipofectamine Stem Cell transfection reagent is more effective than FuGENE in obtaining i PSCs under the conditions tested.



Efficient human iPS cell derivation by a non-integrating plasmid from blood cells with unique epigenetic and gene expression signatures

Using an improved EBNA1/OriP plasmid expressing 5 reprogramming factors, a facile method of generating integration-free human iPSCs from blood MNCs will accelerate their use in both research and future clinical applications.

Generation of Induced Pluripotent Stem Cells from CD34+ Cells across Blood Drawn from Multiple Donors with Non-Integrating Episomal Vectors

This is the first study whereby iPSCs are derived in parallel from multiple donors using episomal, non-integrating, oriP/EBNA1-based plasmids from freshly drawn blood and it is shown that proliferation and cell identity play a role in the number of iPSC per input cell number.

Isolation of human iPS cells using EOS lentiviral vectors to select for pluripotency

The first selection system for the isolation of human iPS cells is reported, and the EOS (Early Transposon promoter and Oct-4 (Pou5f1 and Sox2 enhancers) lentiviral vector is developed to specifically express in mouse and human embryonic stem cells but not in primary fibroblasts.

Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences

Results demonstrate that reprograming human somatic cells does not require genomic integration or the continued presence of exogenous reprogramming factors and removes one obstacle to the clinical application of human iPS cells.

Efficient Feeder-Free Episomal Reprogramming with Small Molecules

Improvements in episomal reprogramming efficiency are greatly improved and enabled the routine derivation of footprint-free human iPSCs from skin fibroblasts, adipose tissue-derived cells and cord blood cells.

Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes

It is shown that reprograming of juvenile human primary keratinocytes by retroviral transduction with OCT4, SOX2, KLF4 and c-MYC is at least 100-fold more efficient and twofold faster compared with reprogramming of human fibroblasts.

A more efficient method to generate integration-free human iPS cells

A simple method is reported, using p53 suppression and nontransforming L-Myc, to generate human induced pluripotent stem cells (iPSCs) with episomal plasmid vectors, which may provide iPSCs suitable for autologous and allologous stem-cell therapy in the future.

Generation of induced pluripotent stem cells from human blood.

The ability to reprogram cells from human blood will allow the generation of patient-specific stem cells for diseases in which the disease-causing somatic mutations are restricted to cells of the hematopoietic lineage.

Improved Efficiency and Pace of Generating Induced Pluripotent Stem Cells from Human Adult and Fetal Fibroblasts

Adding SV40 large T antigen (T) to either set of the four reprogramming genes previously used enhanced the efficiency by 23–70‐fold from both human adult and fetal fibroblasts, and succeeded in replacing MEFs with immortalized human feeder cells that were previously established for optimal hES cell growth.