Evaluation Method Of Vitamin Demand In Human Body By Lymphocyte Culture And Its Application

Vitamin is the essential material to keep human growth and metabolism.Health can not be maintained once deficiency occurred.So it's important to evaluate vitamin status accurately.Human demand for vitamin influenced by many factors,so conventional testing can not reflect the individual needs completely.In this paper,cell culture technology was used,to culture lymphocytes in series medium which containing various nutrients.The difference of cell proliferation capability between different medium was be compared,to evaluate individual's nutritional requirements.Objective:Establish the method of evaluate the individual's six viatmins demand by cell culture technology,and to study the effectiveness of this method.Methods:Prepare series of medium,including completely medium,six nutrients deficiency medium(vitamin B1,B2,B12,Folate,Pantothenic Acid and Biotin).Isolate human lymphocytes from blood,inoculate in medium.After 72 hours incubation,cell division and growth is quantified by tritiated thymidine incorporation assay.Experimental conditions,including PHA concentration in medium,cell inoculate volumn,incubation time,isotope labeling time,the concentration of the isotope were optimized,to make the results stable and reliable.After feasibility experiment,119 volunteers were tested;subjects with abnormal results were treated with oral vitamin supplementation,to evaluate the effectiveness of detection.Results:After optimization of the experimental conditions,four volumteer were tested for four times using completely medium and six nutrients deficiency medium.Results shows that coefficient of variation of four repeating was basically not above 16%for each nutrient;significant differences of R value occurred between volumteers;119 person were tested,22 subjects with abnormal results were treated with oral nutrient supplementation for one month.Then secondly testing was performed.Results shows that R value of all six vitamins increased from 15%to 190%;20 person-time symptoms were improved after supplementation,effective ratio was 83.3%.Conclusions:1.Established the method of evaluate the individual's six vitamins demand by cell culture technology;2.The effectiveness of the method was assessed and confirmed;R value has been increased after vitamin supplementation to those individual with lower R value.