Long-term results of transplanted insulin producing cells differentiated from human adipose derived stem cells in mice

Abstract

Background & Aim We have reported the effective 3D-culture system to differentiate insulin producing cell (IPC) from human adipose derived stem cell (hADSC) recombinant peptide micro-pieces (RCP μ-pieces, FUJIFILM, Tokyo, Japan) (Ikemoto T, et al. Pancreas. 2018, Ikemoto T, et al. Sci Rep 2019, Wada Y, et al. Sci Rep 2019). Although a successful short-term result was confirmed within 30 days with a single administration of IPCs, a long-term fate of IPCs remains unclear. Thus, the aim of this study is to clarify the long-term result of transplanted IPCs in mice. Methods, Results & Conclusion Methods IPCs were generated from hADSCs, which were isolated from patient's subcutaneous adipose, with our original 2-steop protocol which was a xeno-antien free, 3D-culture system as mentioned above (Pancreas. 2018, Sci Rep 2019). BALB/c nu-nu mice were induced to a diabetic state with 200mg/kg streptozotocin for the recipients. 2.0 × 106 IPCs were implanted to these mice under the kidney capsule (Tx group, n=4), same quantity normal saline were administered as control (Sham group, n=4). Non-fasting blood glucose and body weight were measured up to 180 days. Then transplanted mice were sacrificed for immunohistochemical evaluations. Results In Tx group, blood glucose levels in all recipients decreased below 200 mg/dl on day 10 and below 150 mg/dl around day 60 (100%), even though all mice in Sham group were dead in 40 days (100%). The normoglycemic state in Tx group continued until 180 days with IPCs single transplantation (Fig 1). No abnormal tumorous lesions or inflammations were observed at the transplanted sites. Transplanted IPCs were strongly stained by anti-insulin antibody, anti-C peptide antibody and anti-human leukocyte antigen class I antibody (Fig 2). Conclusions Our IPCs can be applied to clinical trial as a promising strategy for type 1 DM patients and totally pancreatectomized patients.