Abstract
Peripheral blood is an easily accessible cell resource for reprogramming into pluripotency by episomal vectors. Here, we describe an approach for efficient generation of integration-free induced pluripotent stem cells (iPSCs) under feeder or feeder-free conditions. Additionally, in combination with the CRISPR-Cas9 genome-editing system, we can directly generate edited iPSCs from blood cells. With this protocol, one can easily generate either integration-free iPSCs or genetically edited iPSCs from peripheral blood at high efficiency.
Keywords:
Episomal vectors; Genomic editing; Induced pluripotent stem cells; Peripheral blood; Reprogramming.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
CRISPR-Cas Systems
-
Cell Culture Techniques / methods*
-
Cells, Cultured
-
Cellular Reprogramming / genetics*
-
Cryopreservation / methods
-
Electroporation / instrumentation
-
Electroporation / methods
-
Feeder Cells
-
Flow Cytometry
-
Gene Editing / methods*
-
Genetic Vectors
-
Humans
-
Immunohistochemistry
-
Induced Pluripotent Stem Cells / cytology*
-
Induced Pluripotent Stem Cells / metabolism
-
Karyotyping
-
Kruppel-Like Factor 4
-
Kruppel-Like Transcription Factors / genetics
-
Kruppel-Like Transcription Factors / metabolism
-
Leukocytes, Mononuclear / cytology*
-
Leukocytes, Mononuclear / metabolism
-
Octamer Transcription Factor-3 / genetics
-
Octamer Transcription Factor-3 / metabolism
-
Plasmids / genetics*
-
Plasmids / metabolism
-
Proto-Oncogene Proteins c-myc / genetics
-
Proto-Oncogene Proteins c-myc / metabolism
-
bcl-X Protein / genetics
-
bcl-X Protein / metabolism
Substances
-
BCL2L1 protein, human
-
Kruppel-Like Factor 4
-
Kruppel-Like Transcription Factors
-
MYC protein, human
-
Octamer Transcription Factor-3
-
POU5F1 protein, human
-
Proto-Oncogene Proteins c-myc
-
bcl-X Protein