Method of rapidly inducing large-scale and high-purity mesenchymal stem cells to transdetermine into hematopoietic stem cells
A method of rapidly inducing large-scale and high-purity mesenchymal stem cells to transdetermine into hematopoietic stem cells is provided with the steps of preparing homogeneous medium of mesenchymal stem cells; combining a plurality of small RNA molecules; assembling and transfecting nanoparticles of nucleic acids and polypeptides; inducing and amplifying medium of post-transdetermined hematopoietic stem cells; and activating a plurality of hemopoiesis-related genes.
1. A method of inducing mesenchymal stem cells to transdetermine into hematopoietic stem cells, comprising the steps of:
(a) preparing a homogeneous culture of mesenchymal stem cells in a non-serum high-glucose Dulbecco's modified Eagle's medium (DMEM) medium comprising epidermal growth factor (EGF), fibroblast growth factor-2 (FGF-2), platelet-derived growth factor subunit B (PDGFB), insulin-like growth factor (IGF), and transforming growth factor (TGF)-beta;
(b) providing a mixture of RNA molecules comprising miR-138-1 and a siR-EID1 (small interfering RNA-EP300-interacting inhibitor of differentiation 1);
(c) assembling nanoparticles comprising the mixture of RNA molecules in step (b) and cell penetrating polypeptides;
(d) transfecting the nanoparticles of step (c) into the culture of mesenchymal stem cells; and
(e) inducing and amplifying hematopoietic stem cells from the transfected mesenchymal stem cells in a non-serum high-glucose DMEM medium comprising sonic hedgehog (shh), stem cell factor (SCF), thyroid peroxidase (TPO), fibromyalgia syndrome-like tyrosine kinase 3 ligand (Flt31), delta-like protein 1 (Delta1), insulin-like growth factor-binding protein (IGFBP), angiopoietin, myelin basic protein 4 (MBP4), leukemia inhibitory factor (LIF), and TGF-beta factors to activate a plurality of hemopoiesis-related genes.