Karyotypic abnormalities in human induced pluripotent stem cells and embryonic stem cells

Induced pluripotent stem cell (iPSC)-based technologies offer an unprecedented opportunity to perform highthroughput screening of novel drugs for neurological and neurodegenerative diseases. Such screenings require a robust and scalable method for generating large numbers of mature, differentiated neuronal cells. Currently available methods based on differentiation of embryoid bodies (EBs) or directed differentiation of adherent culture systems are either expensive or are not scalable. We developed a protocol for large-scale generation of neuronal stem cells (NSCs)/early neural progenitor cells (eNPCs) and their differentiation into neurons. Our scalable protocol allows robust and cost-effective generation of NSCs/eNPCs from iPSCs. Following culture in neurobasal medium supplemented with B27 and BDNF, NSCs/ eNPCs differentiate predominantly into vesicular glutamate transporter 1 (VGLUT1) positive neurons. Targeted mass spectrometry analysis demonstrates that iPSC-derived neurons express ligand-gated channels and other synaptic proteins and whole-cell patch-clamp experiments indicate that these channels are functional. The robust and cost-effective differentiation protocol described here for large-scale generation of NSCs/eNPCs and their differentiation into neurons paves the way for automated high-throughput screening of drugs for neurological and neurodegenerative diseases.

show abstract

“…27 We transfer one iPSC colony from feeder-based culture into a 6-well matrigel-coated plate as follows.…”

Section: Generation and Passaging Of Feeder-free Ipsc Culturesmentioning

confidence: 99%

Large-scale generation of human iPSC-derived neural stem cells/early neural progenitor cells and their neuronal differentiation

et al. 2014

View full text Add to dashboard Cite

show abstract

“…The task is to devise strategies to monitor and evaluate tolerable levels of genetic change and to evaluate the consequences. Numerous studies have compared mutation rates in the original somatic cells and derived iPSCs to analyze at which stage reprogramming affects genomic stability the most (65,66). The major sources of mutations are carryover aberrations from the original cell source, mutations acquired during cell reprogramming, insertional mutagenesis due to the transgenes used for reprogramming, and passage in cell culture (67,68).…”

Section: Challenges To Be Addressed In Preclinical Studiesmentioning

confidence: 99%

Preclinical Studies for Induced Pluripotent Stem Cell-based Therapeutics

Harding

Mirochnitchenko

2014

Journal of Biological Chemistry

View full text Add to dashboard Cite

Induced pluripotent stem cells (iPSCs) and their differentiated derivatives can potentially be applied to cell-based therapy for human diseases. The properties of iPSCs are being studied intensively both to understand the basic biology of pluripotency and cellular differentiation and to solve problems associated with therapeutic applications. Examples of specific preclinical applications summarized briefly in this minireview include the use of iPSCs to treat diseases of the liver, nervous system, eye, and heart and metabolic conditions such as diabetes. Early stage studies illustrate the potential of iPSC-derived cells and have identified several challenges that must be addressed before moving to clinical trials. These include rigorous quality control and efficient production of required cell populations, improvement of cell survival and engraftment, and development of technologies to monitor transplanted cell behavior for extended periods of time. Problems related to immune rejection, genetic instability, and tumorigenicity must be solved. Testing the efficacy of iPSC-based therapies requires further improvement of animal models precisely recapitulating human disease conditions. The breakthrough discovery that specific sets of transcription factors can reprogram cell fate and generate induced pluripotent stem cells (iPSCs) 2 from various cell types has opened many new possibilities for research on cell states, differentiation, pluripotency, and general cell identity but, most importantly, has catalyzed the development of a whole new field of regenerative medicine (1). The field is still in a relatively early stage regarding a clear understanding of underlying developmental processes, cell behavior, and biological effects after cellgrafting experiments. The use of iPSCs and their products for human applications poses many new challenges from the experimental and regulatory points of view due to the unique properties of the cells and novel mechanism of their action.

show abstract

“…Similarly, changes in cell surface proteins due to genetic mutations could also induce an immune response. In addition, in vitro culture itself has been shown to result in genetic instability in PSCs, most commonly chromosomal amplification, including copy number variation (33)(34)(35). These genetic abnormalities could result in not only immunogenicity but also carcinogenicity.…”

Section: Potential Causes Of Ipsc Immunogenicitymentioning

confidence: 99%

The Potential for Immunogenicity of Autologous Induced Pluripotent Stem Cell-derived Therapies

Scheiner

Talib

Feigal

2014

Journal of Biological Chemistry

View full text Add to dashboard Cite

Induced pluripotent stem cell (iPSC) technology offers the promise of immune-matched cell therapies for a wide range of diseases and injuries. It is generally assumed that cells derived from autologous iPSCs will be immune-privileged. However, there are reasons to question this assumption, including recent studies that have tested iPSC immunogenicity in various ways with conflicting results. Understanding the risk of an immune response and developing strategies to minimize it will be important steps before clinical testing. Here, we review the evidence for autologous iPSC immunogenicity, its potential causes, and approaches for assessment and mitigation.

show abstract

Karyotypic abnormalities in human induced pluripotent stem cells and embryonic stem cells

Cited by 308 publications

References 17 publications

Large-scale generation of human iPSC-derived neural stem cells/early neural progenitor cells and their neuronal differentiation

Large-scale generation of human iPSC-derived neural stem cells/early neural progenitor cells and their neuronal differentiation

Preclinical Studies for Induced Pluripotent Stem Cell-based Therapeutics

The Potential for Immunogenicity of Autologous Induced Pluripotent Stem Cell-derived Therapies

Contact Info

Product

Resources

About