ABSTRACT
Providing fetal calf serum [FCS] alternatives as cell culture supplements is an important field of research to compensate for the FCS supply shortage.This study focused on preparation of fetal calf serum alternatives and their effects on growth and secretion of hybridoma cell lines.Outdated human platelet units undergo extraction for its growth factors to be obtained. Human AB blood group plasma was also converted to serum and its growth effect was compared to FCS, hypoxanthine-thymidine [HT] and RPMI1640 as cell culture media and supplement. Cell growth indices were preliminary counting of cells, confluency as surface area of plates filled with cells, and titration of monoclonal anti-A and anti-B blood group antibodies collected from cultured mouse hybridoma cells. Statistical analysis including one sample t-test, logarithmic multiple regression curve fit, and factor analysis was done by SPSS v12 software. The four nutritional supplements of [1] human serum AB [AB], [2] human platelet extract [PLT], [3] equal mixture of AB and PLT [ABP], and [4] fetal calf serum as cell culture were examined on mouse hybridoma anti-A and anti-B monoclonal antibody producer cell lines for cell growth indices and compared with the same indices on RPMI1640 media. The growth-stimulating effects in descending order of values were [1] ABP5%, [2] FCS10%, [3] ABP10%, [4] AB5%, [5] AB10%, [6] PLT5%, [7] ABP20%, [8] PLT10%, [9] PLT20%, and [10]HT; but AB20% inhibited growth of mentioned hybridoma cell lines. The titer of anti-A and anti-B monoclonal antibodies produced by cultured hybridoma on 5 and 10 percent concentration of AB, PLT and ABP compared to FCS5-10% at descending order were [1] PLT5%, [2] PLT10%, ABP5%, ABP10%, AB10%, and [3] AB5%. In general FCS had the following effects on curves of cell growth: [1] the highest increase on slope of multiplication [ascending] phase, [2] the highest increase on slope of death [descending] phase, and [3] the lowest duration of stationary phase. Then, FCS can be appropriate for growth of cells at initial low cell count. Human serum AB, human platelet extract, and equal mixture of both at optimum concentrations [these supplements at high concentrations killed cells] compared to FCS showed [1] decreased slope of multiplication phase, [2] decreased slope of death phase, and [3] increased duration of stationary phase. Thus, AB and PLT may be suitable for continuous cell culture systems in which cell survival during longer times is required. Factor analysis was introduced as a model to evaluate kinetics of cell growth at different supplements
ABSTRACT
One of the natural products in plants is essential oil. Their odor, components and properties are depended on factors such as genetics, climate, habitat and so on. In plant families, Apiaceae is rich in this biochemical group which has different effects in traditional and novel medicine. Comparison of essential oil of two endemic plant of Iran from genus Zeravschania. Zeravschania membranacea [Boiss.] M. Pimenov was collected from Zanjan province in July 2003. The essential oil of powdered aerial parts of plant was prepared by hydro distillation which was about 0.1% [v/w] with yellow color and an intense odor. The essential oil was analyzed with GC/MS on DB-1 column. Thirty five out of forty eamponents, comprising 83.25% of total oil were identified. 44.64% were mono terpenes [15.19% oxygenated, 29.25% hydrocarbons], 7% sesquiterpenes, 7.8% phenyl propanoids and 22.81% nonterpenic compounds. The major components of the above fragments were alpha-Terpenyl acetate [6.6%], Carvacrol [5.95%], Thymol [4.76%], alpha-Ocimene Y [4.08%], beta-Bisabolen [3.24%], Trans-methyl iso Eugenol [7.74%], Thymohydroquinone dimethyl ether [16.7%]. Because of two different habitats and climates for growing Z. membranacea [sandy hill] and Z. pastinacifolia [rocky and scree steep slopes], there are some difference between content of essential oil in them