- The Generation of Human Induced Pluripotent Stem Cells from Blood Cells: An Efficient Protocol Using Serial Plating of Reprogrammed Cells by Centrifugation.
The Generation of Human Induced Pluripotent Stem Cells from Blood Cells: An Efficient Protocol Using Serial Plating of Reprogrammed Cells by Centrifugation.
Stem cells international (2016-09-01)
Youngkyun Kim, Yeri Alice Rim, Hyoju Yi, Narae Park, Sung-Hwan Park, Ji Hyeon Ju
PMID27579041
ABSTRACT
Human induced pluripotent stem cells (hiPSCs) have demonstrated great potential for differentiation into diverse tissues. We report a straightforward and highly efficient method for the generation of iPSCs from PBMCs. By plating the cells serially to a newly coated plate by centrifugation, this protocol provides multiple healthy iPSC colonies even from a small number of PBMCs. The generated iPSCs expressed pluripotent markers and differentiated into all three germ layer lineages. The protocol can also be used with umbilical cord blood mononuclear cells (CBMCs). In this study, we present a simple and efficient protocol that improved the yield of iPSCs from floating cells such as PBMCs and CBMCs by serial plating and centrifugation.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Alkaline Phosphatase Detection Kit, This Alkaline Phosphatase Detection Kit is a specific & sensitive tool for the phenotypic assessment of Embryonic Stem (ES) cell differentiation by the determination of AP activity.
Sigma-Aldrich
Anti-Stage-Specific Embryonic Antigen-4 Antibody, clone MC-813-70, clone MC-813-70, Chemicon®, from mouse