Abstract
Much investigation is needed to understand the underlying molecular mechanisms of iPSC reprogramming and to improve this technology. Lentivirus-mediated iPSC reprogramming remains the most effective method to study human pluripotency reprogramming. iPSC production is more efficient and consistent in the chemically defined medium. Fibroblasts are the most common starting cells for iPSC generation. Here, we provide a detailed protocol for iPSC generation from human fibroblasts using the GFP-expressing lentiviral vectors in the chemically defined medium.
Keywords:
Chemically defined medium; Co-expression of GFP; Fibroblasts; Human iPSC; Human induced pluripotent stem cells; Lentiviral vectors; Pluripotent reprogramming.
Publication types
-
Research Support, N.I.H., Extramural
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Cell Differentiation / genetics*
-
Cells, Cultured
-
Cellular Reprogramming / genetics*
-
Cryopreservation / methods
-
Culture Media / chemistry*
-
Fibroblasts / cytology
-
Fibroblasts / metabolism
-
Flow Cytometry
-
Gene Silencing
-
Genetic Vectors
-
Green Fluorescent Proteins / genetics
-
Green Fluorescent Proteins / metabolism
-
HEK293 Cells
-
HeLa Cells
-
Humans
-
Induced Pluripotent Stem Cells / cytology*
-
Induced Pluripotent Stem Cells / metabolism
-
Kruppel-Like Factor 4
-
Kruppel-Like Transcription Factors / genetics
-
Kruppel-Like Transcription Factors / metabolism
-
Lentivirus / genetics*
-
Lentivirus / metabolism
-
Octamer Transcription Factor-3 / genetics
-
Octamer Transcription Factor-3 / metabolism
-
SOXB1 Transcription Factors / genetics
-
SOXB1 Transcription Factors / metabolism
-
Transcription Factors / genetics
-
Transcription Factors / metabolism*
Substances
-
Culture Media
-
Kruppel-Like Factor 4
-
Kruppel-Like Transcription Factors
-
Octamer Transcription Factor-3
-
POU5F1 protein, human
-
SOX2 protein, human
-
SOXB1 Transcription Factors
-
Transcription Factors
-
Green Fluorescent Proteins