Mesenchymal stromal cells as a reprogramming source for iPSC induction

A method for generating induced mesenchymal stromal cells (iMSCs) comprising:

• obtaining induced pluripotent stem cells (iPSCs) from reprogrammed human fibroblasts (iPSC_Fb) and human primary mesenchymal stromal cells (MSCs);
• culturing the iPSCs with a first medium for about 6 days;
• replacing the first medium with a second medium and culturing the cells with the second medium for about 6 days;
• trypsinizing the cultured cells;
• seeding the trypsinized cells on a coated or non-coated tissue culture with a third medium for about 10 days;
• and
• replacing the third medium with a fourth medium and culturing the seeded cells for 10 days,
• wherein the human primary MSCs are bone marrow derived (BM MSCs),
• wherein a reprogramming of MSCs comprises nonintegrating Sendai virus vector transduction with the addition of Yamanaka transcriptional factors,
• wherein the first medium comprises:
• a Dulbecco's Modified Eagle Medium:
• Nutrient Mixture F-12 (DMEM/F12);
• a knockout serum;
• and
• a transforming growth factor (TGF) beta and anaplastic lymphoma kinase (ALK) inhibitor,
• wherein the second medium comprises:
• DMEM/F12;
• a TGF beta and ALK inhibitor;
• and
• a knockout serum or an Insulin-Transferrin-Selenium (ITS-G) solution,
• wherein the third medium comprises:
• DMEM/F12;
• a knockout serum;
• and
• a basic fibroblast growth factor (bFGF) and an epidermal growth factor (EGF)
• wherein the fourth medium comprises:
• Dulbeccos' Modified Eagle medium-Low Glucose (DMEM-LG) medium and is optionally supplemented with fetal bovine serum (FBS),
• wherein media were changed every two days.