Induction of pluripotent stem cells from fibroblast cultures

  title={Induction of pluripotent stem cells from fibroblast cultures},
  author={Kazutoshi Takahashi and Keisuke Okita and Masato Nakagawa and Shinya Yamanaka},
  journal={Nature Protocols},
Clinical application of embryonic stem (ES) cells faces difficulties regarding use of embryos, as well as tissue rejection after implantation. One way to circumvent these issues is to generate pluripotent stem cells directly from somatic cells. Somatic cells can be reprogrammed to an embryonic-like state by the injection of a nucleus into an enucleated oocyte or by fusion with ES cells. However, little is known about the mechanisms underlying these processes. We have recently shown that the… 

Human iPS cell derivation/reprogramming.

This unit describes a protocol for deriving induced pluripotent stem cells from human fibroblast cells, which share similarities with hES cells including the expression of pluripotency genes, and differentiation as embryoid bodies in vitro into three germ layers (EB) and in vivo as teratomas.

Generation of induced pluripotent mouse stem cells in an indirect co-culture system.

Induced pluripotent stem cells induced by this microporous poly-membrane-based indirect contact co-culture system with mouse embryonic fibroblasts had a reprogramming efficiency and time similar to those induced using traditional methods.

Late Passage Human Fibroblasts Induced to Pluripotency Are Capable of Directed Neuronal Differentiation

Successful reprogramming of human fibroblasts after more than 20 passages in vitro, to a pluripotent state with four transcription factors: Oct4, Sox2, Klf4, and c-Myc is reported.

Generation and characterization of human induced pluripotent stem cells.

These protocols include excellent procedures for passaging and cryopreservation of human iPS cells established by ES cell researchers, which result in an easy way to culture and evaluate the qualities of the generated i PS cells.

Cell reprogramming for the creation of patient-specific pluripotent stem cells by defined factors

In summary, the recent progress in the study of cell reprogramming for the creation of patientspecific pluripotent stem cells, some existing problems, and research perspectives were suggested.

Methods for Inducing Pluripotency

This chapter reviews the scientific foundation that has led to the ability to create iPS cells and the current methods used to make them, as well as the studies that have been conducted to help decipher the molecular pathways involved.

iPS cells produce viable mice through tetraploid complementation

The generation of several iPS cell lines that are capable of generating viable, fertile live-born progeny by tetraploid complementation and confirm that iPS cells can attain true pluripotency that is similar to that of ES cells.

[Induction and characterization of induced pluripotent stem (iPS) cells: a review].

The somatic cells can be induced into ES-like stem cells when retrovirally infected the defined transcription factors including Oct4, Sox2, Klf4 and c-Myc, and to generate iPS cells do need the defined culture procedures.

Induced Pluripotent Stem Cells: A New Approach for Physiological Research

The state of the art in the field of iPS cell induction by cell fusion or defined factors is discussed, techniques to derive pluripotent cells from somatic sources are introduced and discussed, as well as some biological factors that influence the generation ofiPS cells.



How is pluripotency determined and maintained?

These studies show that ES cells continue to self-renew because of a self-organizing network of transcription factors that prevents their differentiation and promotes their proliferation, and because of epigenetic processes that might be under the control of the pluripotent transcription factor network.

Generation of germline-competent induced pluripotent stem cells

iPS cells competent for germline chimaeras can be obtained from fibroblasts, but retroviral introduction of c-Myc should be avoided for clinical application.

Developmental reprogramming after chromosome transfer into mitotic mouse zygotes

It is reported that, unlike interphase zygote, mouse zygotes temporarily arrested in mitosis can support somatic cell reprogramming, the production of embryonic stem cell lines and the full-term development of cloned animals.

Nanog promotes transfer of pluripotency after cell fusion

It is reported that in fusions between ES cells and neural stem (NS) cells, increased levels of Nanog stimulate pluripotent gene activation from the somatic cell genome and enable an up to 200-fold increase in the recovery of hybrid colonies, all of which show ES cell characteristics.

Self-renewal of teratocarcinoma and embryonic stem cells

Recent progress in mouse embryonic stem cells self-renewal is reviewed, this to the biology of teratocarcinomas and testable hypotheses to expose the mechanics of ES cell self-Renewal are offered.

Plat-E: an efficient and stable system for transient packaging of retroviruses

A potent retrovirus packaging cell line named Platinum-E (Plat-E) was generated based on the 293T cell line and the probability of generating the replication competent retroviruses (RCR) by recombination can virtually be ruled out.