The Differences in the Characteristics of Insulin-producing Cells Using Human Adipose-tissue Derived Mesenchymal Stem Cells from Subcutaneous and Visceral Tissues

Abstract

The aim of this study was to investigate the characteristics of insulin producing cells (IPCs) differentiated from adipose-tissue derived stem cells (ADSCs) isolated from human subcutaneous and visceral adipose tissues and identify ADSCs suitable for differentiation into efficient and functional IPCs. Subcutaneous and visceral adipose tissues collected from four (4) patients who underwent digestive surgeries at The Tokushima University (000035546) were included in this study. The insulin secretion of the generated IPCs was investigated using surface markers by: fluorescence activated cell sorting (FACS) analysis; cytokine release; proliferation ability of ADSCs; in vitro (glucose-stimulated insulin secretion: (GSIS) test/in vivo (transplantation into streptozotocin-induced diabetic nude mice). The less fat-related inflammatory cytokines secretions were observed (P < 0.05), and the proliferation ability was higher in the subcutaneous ADSCs (P < 0.05). Insulin expression and GISI were higher in the subcutaneous IPCs (P < 0.01 and P < 0.05, respectively). The hyperglycaemic state of all mice that received IPCs from subcutaneous fat tissue converted into normo-glycaemia in thirty (30) days post-transplantation (4/4,100%). Transplanted IPCs were stained using anti-insulin and anti-human leukocyte antigen antibodies. The IPCs generated from the ADSCs freshly isolated from the human fat tissue had sufficient insulin secreting ability in vitro and in vivo.

Generation of Insulin-Producing Cells from Canine Adipose Tissue-Derived Mesenchymal Stem Cells

“…The study demonstrated that intraperitoneal transplantation of 1.5 × 10 6 BM-MSCs that differentiated into IPCs into diabetic rats led to the improvement of the hyperglycemic states. However, in a recent report on the transplantation of IPCs generated from human AT-MSCs with μ -pieces into diabetic mice, 4.8 × 10 7 cells were transplanted under the kidney capsule or intramesentery region and led to normoglycemic states [ 44 , 45 ]. One of these studies also calculated the cell number of generated IPCs and found that 1.8 × 10 4 IPCs were obtained from 5 × 10 5 AT-MSCs, indicating that 3.6% of the initial AT-MSCs differentiated into IPCs; thus, approximately 1.7 × 10 6 IPCs are needed per mouse to gain the normoglycemic states in STZ-induced diabetic mice [ 45 ].…”

“…However, in a recent report on the transplantation of IPCs generated from human AT-MSCs with μ -pieces into diabetic mice, 4.8 × 10 7 cells were transplanted under the kidney capsule or intramesentery region and led to normoglycemic states [ 44 , 45 ]. One of these studies also calculated the cell number of generated IPCs and found that 1.8 × 10 4 IPCs were obtained from 5 × 10 5 AT-MSCs, indicating that 3.6% of the initial AT-MSCs differentiated into IPCs; thus, approximately 1.7 × 10 6 IPCs are needed per mouse to gain the normoglycemic states in STZ-induced diabetic mice [ 45 ]. In our study, we did not calculate the differentiation ratio of IPCs, but the differentiation protocol referred to in our study reported approximately that 2.5% of human BM-MSCs differentiate into insulin-positive cells under a 2D conventional culture system [ 17 ].…”

“…In our study, we collected adipose tissue from falciform ligament fat and isolated AT-MSCs. A previous study on the insulin secretion capacity of IPCs generated from human AT-MSCs reported that IPCs generated from visceral AT-MSCs showed poor differentiation and insulin secretion compared with those from subcutaneous AT-MSCs [ 45 ]. One reason why visceral AT-MSCs have been not suitable for differentiation into IPCs is that visceral AT-MSCs secrete more inflammatory cytokines such as IL-6 and TNF- α [ 46 ].…”

“…The study also indicated that IPCs generated from visceral AT-MSCs were inferior regarding glucose-stimulated insulin secretion. A stimulation index (SI), a parameter that reflects the insulin releasing function in response to glucose levels, of less than 3 does not normalize blood glucose levels after transplantation [ 45 ]. Our generated IPCs from canine visceral AT-MSCs showed increasing insulin secretion in response to high glucose levels, but the SI of our generated IPCs in vitro was only approximately 1.4 (calculated as the level of insulin concentration under high glucose condition (25 mM) divided by that under low glucose condition (5.5 mM)).…”

Viability, Yield and Expansion Capability of Feline MSCs Obtained from Subcutaneous and Reproductive Organ Adipose Depots

“…MSCs are becoming increasingly relevant to clinical care [7] but the outcomes associated with MSC treatment have shown great inconsistency. Some of the outcome variability may be due to the altered characteristics associated with the cells, based on the site of isolation of the initial tissue [13,27]. Other factors responsible for inducing variability may include the methods of isolation, the amount of tissue harvested, and the passage number.…”