| At present, aging is becoming an important social problem. The exploring for effective anti-aging measures and amendatory survive-quality steps turn into the urgent affairs. Recent research has revealed that melatonin is known as a stringent biological modulator of mood, sleep, retinal physiology, sexual behavior, seasonal reproductive physiology and behavior, circadian rhythms, and immunologic function. Morever, there is experimental evidence that reflects the influence of melatonin on aging and age-related process and disease states for its ability of neutralizing free radicals. Therefore, aging and aging-depentent disease are amenable to treatment with melatonin. However, exogenous melatonin or melatonin -like drugs are still under investigation in a wide range of biomedical research areas to definite its negative or side-effects before it is applied in clinic widely and safely. It is likely that microencapsulation of pinealocytes is expected to play a vital role in future anti-aging process. Microencapsulated pinealocytes will exert its effects of anti-aging process by continuously melatonin secreting after grafting on aging animals, while, this technique refrain from indefinite curative effect and side effect of exogenouly administered melatonin and immune rejection of host animal. In this study, we explored the growth, morphology, proliferation and the biological characteristics of cultured and encapsulated rat pinealocytes in vitro, and examined the correction for serum concentration of model rats with microencapsulation pinealocytes transplantatiorn. Our approach is therefore to lay a foundation that serve as the basis for the development of tissue engineering method to confront aging process.Materials and Methods (1) . Setting up pinealocytes culture system in vitro: The pineal body ofneonatal rats was removed by microsurgical technique. The pinealocytes were then isolated by digesting method with trypsin and collagenase. Cells were cultured in DMEM culture medium added with 10% TBS. The growth and morphologic characteristics were observed by the inverted phase contract microscope.(2) . The proliferation, differentiation, functional express and thebiological characteristics of pinealocytes at different cultured time were measured by MTT, BrdU, 5-HT immunocytochemical assay (SABC), HPLC,transmission electron microsope and RT-PCR. methods. The best time of proliferation and differentiation states of cultured pinealocytes were explored.(3) . APA pinealocytes microcapsules of 0.4-0.7mm diamete wereobtained by co-axis air-jet system and cultured in vitro, and the cell viability, the function and the immunogenicity of pinealocytes microencapsulation were investigated.(4) . The pinealocytes came from cultured or directly digested wereencapsuled respectively and their functional differences were compared. The tolerance, adaptability and activity of two kinds of encapsules were screened so that the favorable pinealocytes can be used for encapsulation and transplantation.(5) . To establish model of pinealectomized rats and detect dynamicchange of serum MT concentration of model animals. The best time of encapsulation transplantation was evaluated and cleared cut. ( 6 ) . To transplant and retrieve the pinealocytes microencapsulation .The morphologic and structured changes of retrieve capsules were analysed, furthermore, the function, immunogenicity and biocompatility of pinealocytes microencapsulation in vivo were assessed by studying studying dynamic of serum MT concentration of model animals after transplantation.Results(1) . Pinealocytes grew vigorously under the ordinary culture condition(DMEM-F12, contain 10%FBS) after digesting .The primary cultured pinealocytes maintained well survive state for nearly 60 days.(2) . The isolated cells were identified to be pinealocytes. The time ofcellular group duplication was the ninth days and reached it' s peak at the tenth days.The secretion function of pinealocytes i...
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