Generation of integration-free human induced pluripotent stem cells from postnatal blood mononuclear cells by plasmid vector expression.
Several human postnatal somatic cell types have been successfully reprogrammed to induced pluripotent stem cells (iPSCs). Blood mononuclear cells (MNCs) offer several advantages compared with other cell types. They are easily isolated from umbilical cord blood (CB) or adult peripheral blood (PB), and can be used fresh or after freezing. A short culture allows for more efficient reprogramming, with iPSC colonies forming from blood MNCs in 14 d, compared with 28 d for age-matched fibroblastic cells. The advantages of briefly cultured blood MNCs may be due to favorable epigenetic profiles and gene expression patterns. Blood cells from adults, especially nonlymphoid cells that are replenished frequently from intermittently activated blood stem cells, are short-lived in vivo and may contain less somatic mutations than skin fibroblasts, which are more exposed to environmental mutagens over time. We describe here a detailed, validated protocol for effective generation of integration-free human iPSCs from blood MNCs by plasmid vectors.
Stem Cell Program, Institute for Cell Engineering, Division of Hematology, Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Sarah N
SN
Dowey
Xiaosong
X
Huang
Bin-Kuan
BK
Chou
Zhaohui
Z
Ye
Linzhao
L
Cheng
D018929:Cell Culture Techniques; D005822:Genetic Vectors; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007963:Leukocytes, Mononuclear / Q000166:cytology*; D010957:Plasmids / Q000235:genetics; D014162:Transfection