The Immunoregulation Of Adipose Derived Stem Cells Transplantation On Food Allergy Of Balb/c Immature Mice

Objective: 1. To separate Adipose Derived Stem Cells from Balb/c mice and do identification and cultivation successfully. 2. To build food allergy model of Balb/c immature mice with ovalbumin(OVA), and to detect the serum levels of cytokines by the Luminex liquid chip technology. 3. To explore the immunoregulation effect of Adipose Derived Stem Cells(ADSC) transplantation on allergic immature mice before and after sensitized with ovalbumin(OVA).Method: SPF Balb/c male adult mice were killed by cervical dislocation executed, take adipose tissue from epididymis and its surrounding areas on super clean bench, cut adipose tissue into pieces with eye scissors, then add I collagenase for digestion. Filter to remove undigested tissue, inoculate cells after centrifugal, then cultivate in cell culture box. High purity of ADSC was obtained after three to four generations. Induce ADSC into fat, osteogenesis for identification and detect surface antibody of CD44, CD29, then cryopreserve for transplantation after identification. Thirty two 3-week old female mice were randomly divided into blank control group, allergic model group, the group treated and prevented with ADSC, each group eight. The food allergy model of Balb/c immature mice were built intraperitoneally with OVA; The ADSC transplantation, 1×106 stem cells for each mouse of the preventive and the treatment groups, was carried Intraperito- neally out at the first day, the fifteenth day sensitized respectively; replaced ADSC with equal PBS for the blank control group and the allergic model group. The allergic symptoms of all groups were scored. All mice were executed after the last inspire sensitization and 0.5cm length jejunum were taken for HEstaining. Observe the pathological difference of vary groups, and collect blood by picking eyeball before execution. Level of the allergy-related inflammatary cytokines in serum were detected for the all groups with Luminex technology.Result: 1. In vitro, Adipose Derived Stem Cells have been successfully separated from Balb/c mice adipose. ADSC can appear fat drops with fat inducers for 2 weeks, and it can appear red by Oil Red O staining; ADSC can appear calcium nodules after osteogenesis inducers for three weeks, then can appear red alizarin red staining. ADSC positive express CD44(98.92%), CD29(99.78%). 2. We build food allergy model of Balb/c successfully. Compared with the blank control group, model group showed obvious symptoms. Scoring average(3.13 ± 0.64) of allergic symptoms in model mice is significantly higher than the blank control group(0.38 ± 0.52)(P <0.05). Pathological appearance of intestinal villus is edema and infiltration of inflammatory cells. IFN-γ, the characteristic cytokine of Th1 in serum, is decreased(P<0.05), while IL-4, the characteristic cytokine of Th2 is increased(P< 0.05). Levels of Other cytokines of allergic model group IL-5, IL-6, IL-17 A, IL-22, IL-23, MCP-1, are elevated compared with control group, and the changes were statistically significant(P<0.05). IL-10 and IL-13 are increased, but no statistical significance(P > 0.05). 3. Compared with allergic model group(3.13 ± 0.64), the allergic symptoms scores are obviously reduced in the treatment group(1.75±0.71, P<0.05). The allergic pathological damages in this group are improved. The allergy-related cytokines, such as IL-4, IL-5, IL-6, IL-17 A, IL-22, IL-23, MCP-1 are distinctly lower in the ADSC-treated group than in the food allergy group(P<0.05), but the Th1-related inflammatory cytokine IFN-γ higher in the treated group than that in the food allergy group(P<0.05). ADSC transplantation can play some treating immunoregulation effect in the treatment group with ADSC 4. Allergic symptom scores of the ADSC preventive group is lower than the allergicmodel group(P < 0.05), but is higher than the ADSC treatment group. The pathological appearance of allergy group is remission. Compared with the allergic model group, the levels of cytokines IL-4, IL-5, IL-6, IL-22, IL-23 are statistically significant(P < 0.05), and the change of the others indexes has no statistical significance(P > 0.05). In both the prevented and treated groups, ADSC transplantation can alleviate the symptoms of food allergy mice and play some preventive or treating immunoregulation effect in the treatment group and the preventive group with ADSC, more significant for the treatment group.Conclusion: 1. We can achieve ADSC in vitro by preparation, separation and cultivation of immature rat’s adipose tissue successfully; 2. We can build food allergy model with 3 weeks of Balb/c immature rat; 3. ADSC transplantation can alleviate the food allergy symptoms, improve allergic pathological damage, reduce allergy-related cytokines of Balb/c immature mice and play some preventive or treating immunoregulation effect in the treatment group and the preventive group of ADSC, more significant for the treatment group.