title: GM-CSF and MEF-conditioned media support feeder-free reprogramming of mouse granulocytes to iPS cells.
abstract: Induced pluripotent stem cells (iPSCs) are characterised by their ability to differentiate into any cell type of the body. Accordingly, iPSCs possess immense potential for disease modelling, pharmaceutical screening and autologous cell therapies. The most common source of iPSCs derivation is skin fibroblasts. However, from a clinical point of view, skin fibroblasts may not be ideal, as invasive procedures such as skin biopsies are required for their extraction. Moreover, fibroblasts are highly heterogeneous with a poorly defined developmental pathway, which makes studying reprogramming mechanistics difficult. Granulocytes, on the other hand, are easily obtainable, their developmental pathway has been extensively studied and fluorescence activated cell sorting allows for the isolation of these cells at high purity; thus iPSCs derivation from granulocytes could provide an alternative to fibroblast-derived iPSCs. Previous studies succeeded in producing iPSC colonies from mouse granulocytes but with the use of a mitotically inactivated feeder layer, restricting their use for studying reprogramming mechanistics. As granulocytes display poor survival under culture conditions, we investigated the influence of haematopoietic cytokines to stabilise this cell type in vitro and allow for reprogramming in the absence of a feeder layer. Our results show that treatment with MEF-conditioned media and/or initial exposure to GM-CSF allows for reprogramming of granulocytes under feeder-free conditions. This work can serve as a basis for future work aimed at dissecting the reprogramming mechanism as well as obtaining large numbers of iPSCs from a clinically relevant cell source.
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton 3800, Victoria, Australia; Australian Regenerative Medicine Institute, Monash University, Clayton 3800, Victoria, Australia.
authors.forename: Jaber
authors.initials: J
authors.lastname: Firas
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton 3800, Victoria, Australia; Australian Regenerative Medicine Institute, Monash University, Clayton 3800, Victoria, Australia.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton 3800, Victoria, Australia; Australian Regenerative Medicine Institute, Monash University, Clayton 3800, Victoria, Australia. Electronic address: christian.nefzger@monash.edu.
authors.forename: Christian M
authors.initials: CM
authors.lastname: Nefzger
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton 3800, Victoria, Australia; Australian Regenerative Medicine Institute, Monash University, Clayton 3800, Victoria, Australia. Electronic address: jose.polo@monash.edu.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Polo
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D065150:Cellular Reprogramming / Q000187:drug effects; D017077:Culture Media, Conditioned; D005347:Fibroblasts / Q000187:drug effects; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology*; D006098:Granulocytes / Q000166:cytology* / Q000187:drug effects; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects; D051379:Mice
chemical_list: D017077:Culture Media, Conditioned; D016178:Granulocyte-Macrophage Colony-Stimulating Factor
keywords: Blood; GM-CSF; Granulocytes; Induced pluripotent stem cells; MEF-conditioned media