| Diabetes mellitus(DM)has become a common disease and one of metabolic endocrine diseases in companion animals.Its causes are associated with obesity,immunity,drugs,pancreatitis and other inducing factors.Typical clinical symptoms include polydipsia,polyuria,polyphagia and weight loss.Current treatments include injecting insulin or transplantation of the inducing stem cells into insulin-secreting cells to restore islet function.However,these methods still have the problems of high cost or difficult to prepare on a large scale.Therefore,in the clinical treatment of clinical diabetes in pets,a low-cost,convenient and effective treatment method is urgently needed to solve the current issue.Adipose-derived mesenchymal stem cells(ADSCs)therapy has gradually entered the attention of researchers.ADSCs are derived from mesoderm,and have the potential of high self-renewal and multi-differentiation.At the same time,ADSCs also have the characteristics of low immunogenicity,and are relatively easy be isolated,cultured and amplified.Compared with other cells,ADSCs show strong therapeutic potential in the research of various diseases.Therefore,the emergence of ADSCs has greatly promoted the development of cell transplantation therapy.Although ADSCs have abundant sources and low immunogenicity,there are still many problems,such as cumbersome separation and purification,differences between generations,aging and so on,which will affect the therapeutic effect of cells.Immortalized cells are a kind of stable and homogeneous cells with unlimited proliferation ability,which can be used as ideal seed cells for transplantation therapy.Previously,our research group successfully established an immortalized cell line of ADSCs.Based on this,we established an animal model of diabetes in mice,and transplanted primary and immortalized ADSCs into mice by intravenous injection.Up to now,there are few reports on the therapeutic effects of immortalized cell lines on diseases,and this study aims to explore the therapeutic effects of primary and immortalized ADSCs cell lines on diabetes mellitus,and compare the differences of different cell lines to select suitable cells for subsequent research and application.In this experiment,primary and immortalized ADSCs were resuscitated and subcultured in vitro,and then transplanted into animal models.24 KM mice and 8 dogs were randomly divided into four groups:normal control group,model group,primary ADSCs cell treatment group and immortalized ADSCs cell treatment group.Four Chinese pastoral dogs were randomly divided into groups,and the experiment was conducted after all indexes were normal.Except the control group,the other three experimental groups were fed with high fat and high sugar combined with streptozocin(STZ)to establish DM model,while the mice in the control group were injected with equal dose of normal saline.Mice received cell transplantation on the 0 th,7 th and 14 th day after the establishment of the model,and all the control animals received the same amount of normal saline injection.Another 18 KM mice and 3 dogs were selected for safety evaluation.The blood sugar,biochemical indexes and tissues of experimental animals were detected,and the following results were obtained:(1)Model evaluation:After continuous intraperitoneal injection of STZ,the blood glucose of mice and canine increased,the body weight lost,and the spirit was depressed.The animal with random blood glucose greater than 11.1 mmol/L met the standard of DM,and the pancreatic tissue had obvious injury characteristics,which indicated that DM model was successfully established.The success rate was 86.7%in mice and 100%in dogs.After cell transplantation,the blood sugar of mice decreased,glucose tolerance and insulin tolerance recovered significantly,the biochemical indicators were improved,and tissue damage has been significantly reduced.(2)Efficacy evaluation:By comparing various data,primary ADSCs have significantly better ability to regulate blood glucose than immortalized ADSCs,and have similar effects on tissue structure repair and biochemical indexes.The results showed that although the immortalized ADSCs cell lines had fast proliferation rate and stable batch and could be transmitted indefinitely,their ability to regulate blood glucose was not very good in the treatment of diabetes,their ability to repair tissue damage and regulate biochemical indexes was not different from that of the primary ADSCs,which proved that intravenous infusion of the two cell lines had varying degrees of effectiveness.(3)Safety evaluation:In the safety experiment,livers of mice were obtained by intravenous infusion of 1*106 primary ADSCs and immortalized ADSCs in mice and dogs on the 0th,7th and 14th day Pancreas and groin fat were found to be normal in tissue morphology after implantation,and no obvious abnormality was found after detection of various biochemical indexes of dogs.Both mice and dogs survived safely,with a survival rate of 100%,indicating that both cell lines were safe for intravenous infusion.This study proved that immortalized adipose-derived mesenchymal stem cells and primary adipose-derived mesenchymal stem cells were isolated and cultured,the primary adipose-derived mesenchymal stem cells were more effective in treating DM by intravenous infusion in diabetic model mice,and both cell lines have safe efficacy.This experiment provides a certain experimental basis and application basis for mesenchymal stem cells to treat diabetes. |
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