Effects of postharvest pretreatments and preservative solutions on vase life longevity and flower quality of sweet pea (Lathyrus odoratus L.)

The effects of postharvest pretreatments on vase life, keeping quality and carbohydrate concentrations in cut sweet pea (Lathyrus odoratus L.) flowers were investigated. Compared to the control, all treatments promoted floret quality and extended longevity. The cut flowers held in the solution containing sucrose + 8-hydroxyquinoline (Suc+HQS) was more effective in promoting absorption rate, achieved greater maximum fresh mass, had better water balance for a longer period, extended the vase life (up to 17 d), and delayed degradation of chlorophylls. The same treatment also enhanced the concentration of soluble carbohydrates in the petals and stems and leaf chlorophyll (Chl) content, whereas it was lowest in silver thiosulphate (STS) treatment. However, concentrations of anthocyanin in the petals were higher for treatment with sucrose or STS plus sucrose than in control or STS alone treatments. Our results suggest that pulse treatment with HQS plus sucrose for 12 h is the most effective for improving pigmentation and use as a commercial cut flower preservative solution to delay flower senescence, enhance quality, and prolong the vase life of sweet pea. The results also showed that soluble carbohydrate concentration in petals and stems is an important factor in determining the vase life of sweet pea flowers.     

Vase life of cut rose cultivars ‘Avalanche’ and ‘Fiesta’ as affected by Nano-Silver and S-carvone treatments

Rosa hybrida L. is an important commercial cut flower. The vase life of this flower is usually short due to vascular occlusion. We assessed the effect of Nano-Silver (NS) and S-carvone in prolonging the vase life of R. hybrida L. cv. ‘Avalanche’ and ‘Fiesta’. Hence, an experiment involving the treatment with NS at 0, 50, 100, and 200 mgL^− 1 and S-carvone at 0, 0.25, 0.5, and 0.75 mgL^− 1 with 10 replicates was conducted. Applying NS pulse treatments increased vase life, water uptake rate, and fresh weight and reduced the number of bacteria, water loss, stomatal conductance and transpiration rate. However, S-carvone treatments did not have a positive effect on the vase-life parameters of cut rose flowers. NS pulse treatments increased relative fresh weight (RFW), and water uptake rate (WUR) and decreased water loss (WL) (%) by 10, 89 and 31% for cultivar ‘Avalanche’, compared to the controls, respectively. Application of 200 mgL^− 1 NS led to the highest vase life (18 days) for roses. The results show that NS increased vase life by suppressing stomatal opening, decreasing transpiration from leaves and inhibiting bacterial proliferation.     

γ射线紫外线镰刀菌产生玉米赤霉烯酮毒素的影响

锭刀菌(Fusarium)经γ射线、紫外线辐射诱变后,接种于大米培养基中.经10℃低温产毒培养.采用薄层层析方法定性定量分析培养物中的镀刀菌毒素—玉米赤霉烯酮(zearalenone简称ZEA).结果表明:不同菌株对诱变剂的反应不同.产毒菌株NF5232、NF5946在较大剂量γ射线处理后,ZEA产量显著增加,但其产生的色素量下降.或者用紫外线处理,仍然产生ZEA.不产毒菌株NF6127、NF6123、NP6138和HD—002,经γ射线或紫外线处理后仍不产生ZEA.由此可知镰刀菌产生ZEA毒素的能力是相对稳定的,其产毒机制是由菌株本身的遗传特性决定的,一定剂量的γ射线可使低产毒菌株变为高产毒菌株.     

Ethylene- and dark-induced flower abscission in potted Plectranthus: Sensitivity,prevention by 1-MCP,and expression of ethylene biosynthetic genes

Prevention of ethylene- and shipping-induced flower abscission is necessary to maintain the quality of both cut flowers and potted plants during handling, transport and retail display. The aims of the present work were to determine the sensitivity of Plectranthus cultivars to applied ethylene, to alleviate ethylene- and shipping-induced flower abscission in intact potted plants using 1-methylcyclopropene (1-MCP), and to investigate the possible causes of dark-induced flower abscission. All cultivars were sensitive to ethylene in a concentration-dependent manner, and complete abscission occurred within 24 h with 1 and 2 μl l^− 1 ethylene. Unopened buds were more sensitive to applied ethylene, and exhibited greater abscission than open flowers. Ethylene synthesis remained below detection limits at all time points under control and continuous dark conditions. Dark treatment significantly increased flower abscission in Plectranthus cultivars, and like ethylene-induced flower abscission, this could be prevented by continuous 1-MCP treatment. Gene expression of ethylene biosynthetic enzymes ACS and ACO was examined as possible causes for the accelerated flower abscission observed in plants kept in continuous darkness. Expression patterns of ACS and ACO varied between different cultivars of Plectranthus. In some cases, increased expression of ACS and ACO led to increased flower abscission. Gene expression was higher in open flowers when compared to unopened flowers suggesting a cause for the observed preferential shedding of open flowers in some cultivars. Although the cause of dark-induced abscission in Plectranthus remains elusive, it can be effectively controlled by treatment with 1-MCP.     

Flower model traps reduced thrips infestations on a pepper crop in field

A flower model trap developed by modifying an artificial yellow chrysanthemum flower was more attractive to flower thrips than commercial yellow sticky traps. Installation of these flower model traps (20 traps per 50 m² plot) was reported to reduce seasonal populations of Frankliniella intonsa (Trybom) (Thysanoptera: Thripidae) on strawberry flowers in greenhouses. In this study, we sought to determine if the installation of such flower model traps would reduce thrips populations in a pepper field. The traps were installed at the bottom of the plant canopy at varying densities (0, 5, 10, and 20 traps) in 20 plots (each 3 × 5 m²) using a completely randomized design. Thrips populations on pepper flowers were sampled from 1 to 29 July in 2009. All thrips sampled on the flowers were identified as F. intonsa. A significant effect of treatment and sampling date was found from repeated-measure analysis of variance. The highest density (20 traps per 15 m²) of traps significantly reduced the female and male F. intonsa population compared to the control by 61 and 49%, respectively. However, no difference in immature thrips numbers was found among the treatments. These results indicate that this flower model trap can be a useful tool for the management of flower thrips on field-grown peppers.     

Antimicrobial activities of Asafoetida and Shirazi thyme essential oils improve the vase life of gerbera cut flowers

This study was conducted to evaluate the chemical composition of asafoetida (Ferula assa-foetida) essential oil (FAEO) and Shirazi thyme (Zataria multiflora) EO (ZMEO) and their impact on vase life of gerbera cut flowers (Gerbera jamesonii cv. Rosalyn). Five concentrations of both, ZMEO and FAEO including 0, 100, 200, 300 and 400 mg L^?1 used as continuous vase solution for gerbera cut flowers. EOs used in this study were extracted by hydrodistillation method using Clevenger apparatus. They were analyzed by GC and GC–MS for determination of the active compounds. Thirty five compounds were identified in ZMEO, mainly including thymol (40.1%), p-cymene (15.5%) and carvacrol (6.5%). Also, thirty compounds were identified in FAEO. The main components were trans propenyl sec-butyl disulfide (21.7%), eudesmol (10-epu-γ) (19.2%) and cis propenyl sec-butyl disulfide (10.2%). The results showed that both ZMEO and FAEO at all concentrations could act as an effective antibacterial compounds and this property increased by increasing their concentration. The results of this research showed that ZMEO increased the vase life at all concentrations but high concentrations of FAEO increased mortality percentage and reduced the vase life of cut flowers. The relative fresh weight and vase solution uptake of gerbera cut flowers increased by the applied EOs treatments. ZMEO at 400 mg L^? 1 and FAEO at 300 and 400 mg L^? 1 resulted the least stem color change. Overall, 200 mg L^? 1 ZMEO and 100 mg L^? 1 FAEO were the best treatments for maintenance of gerbera cut flowers quality during vase life.     

Microwave-assisted drying of paclitaxel for removal of residual solvents

In this study, the residual solvent in final purified paclitaxel was effectively removed using microwave-assisted drying. When the sample final purified by silica-HPLC was concentrated using a rotary evaporator, the residual methanol easily met the ICH-specified value (3000 ppm), but methylene chloride did not meet the ICH-specified value (600 ppm). Thus, the efficiency of microwave-assisted drying according to microwave power (100, 200, and 300 W) and drying time was investigated using the sample (methylene chloride conc.: 26,000 ppm, methanol conc.: 50 ppm) concentrated by rotary evaporation. A higher microwave power was effective in removing methylene chloride, and the ICH requirements were met by drying at 300 W for 21 h. In addition, when the sample concentrated by rotary evaporation was vacuum dried (35 °C, 24 h), the concentration of methylene chloride could be reduced to 8500 ppm. When the vacuum-dried sample was subjected to microwave-assisted drying, the ICH requirements could be met by drying for 10 h at 200 W and 8 h at 300 W. The lower the initial concentration of the solvent and the higher the microwave power, the greater the improvement in the efficiency of microwave-assisted drying.     

Effects of antimicrobial activity of silver nanoparticles on in vitro establishment of G × N15 (hybrid of almond × peach) rootstock

In the present investigation were evaluated the antifungal and antibacterial activities of Nano-silver (NS). Two separate experiments were done to evaluate the potential of silver nanoparticles in controlling the contamination of G × N15 micro-propagation. In the first experiment, a factorial experiment based on a completely randomized design with 15 treatments including five different NS concentrations (0, 50, 100, 150 and 200 ppm) and three soaking time of explants (3, 5 and 7 min) with five replications was conducted. In the other experiment, NS was added to Murashige and Skoog (MS) medium with concentrations of 0, 50, 100, 150 and 200 ppm in a completely randomized design. Results showed that the application of 100 and 150 ppm NS both as an immersion and as added directly to the culture medium significantly reduces internal and external contaminations compared with the control group. Using NS in culture medium was more efficient to reduce fungal and bacterial contamination than immersion. High concentrations of NS had an adverse effect on the viability of G × N15 single nodes and this effect was more serious in immersed explants in NS than directly added NS ones regarding the viability of buds and plantlet regeneration. In conclusion, due to high contamination of woody plants especially fruit trees and also adverse environmental effects of mercury chloride, the NS solution can be used as a low risk bactericide in micro-propagation of G × N15 and can be an appropriate alternative to mercury chloride in the future.     

Production of isomaltulose using Erwinia sp. D12 cells: Culture medium optimization and cell immobilization in alginate

Isomaltulose is a structural isomer of sucrose commercially used in food industries. Glucosyltransferase produced by Erwinia sp. D12 catalyses an intramolecular transglucosylation of sucrose giving isomaltulose. An experimental Design and Response Surface Methodology were applied for the optimization of the nutrient concentration in the culture medium for enzyme production in shaken flasks at 200 rpm and 30 °C. A higher production of glucosyltransferase (7.47 Uml⁻¹) was observed in the culture medium containing sugar cane molasses (160 gl⁻¹), bacteriological peptone (20 gl⁻¹) and yeast extract Prodex Lac SD^® (15 gl⁻¹) after 8 h, at 30 °C. The highest production of glucosyltransferase in the 6.6-l bioreactor (14.6 Uml⁻¹) was obtained in the optimized culture medium after 10 h at 26 °C. When Erwinia sp. D12 cells were immobilized in sodium alginate, it was verified that sodium alginate solution A could be substituted by a cheaper one, sodium alginate solution B. Using a 40% cell suspension and 2% sodium alginate solution B for cell immobilization in a packed-bed reactor, 64.1% conversion of sucrose to isomaltulose was obtained. The packed-bed reactor with immobilized cells plus glutaraldehyde and polyethylenimine solutions remained in a pseudo-steady-state for 180 h.     

Low-temperature phosphine fumigation of chilled lettuce under insulated cover for postharvest control of western flower thrips,Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae)

Pallet-scale phosphine fumigations were conducted on pre-chilled iceberg lettuce under an insulated cover to determine efficacy against western flower thrips, Frankliniella occidentalis, and phytotoxicity to lettuce. Vacuum-cooled commercial iceberg lettuce at 3 °C was sealed in a plastic bag, covered with double-bubble foil insulation, and then fumigated with 484 ± 17 ppm phosphine for 18 h under storage at 20 °C. Lettuce temperature increased from 4.5 °C to 7.2 °C during the course of fumigations. The fumigation treatment was replicated three times and achieved complete control of 3453 western flower thrips. Lettuce quality was evaluated 2 weeks after fumigation and the fumigation had no significant negative impact on lettuce quality. This study demonstrates that insulated cover can be used to keep pre-chilled lettuce at low temperature suitable for low-temperature phosphine fumigation to control western flower thrips on harvested lettuce.     

Ultralow oxygen treatment for control of western flower thrips,Frankliniella occidentalis (Thysanoptera: Thripidae), on harvested table grapes

Exported organic table grapes from the U.S. are currently fumigated with methyl bromide to control quarantined pests such as western flower thrips, Frankliniella occidentalis (Pergande). An alternative treatment which is compatible with organic products is needed. In this study, controlled atmosphere with ultralow oxygen (ULO) treatment was studied to control thrips on harvested table grapes. ULO treatments with < 0.01 ppm oxygen at 3.3 °C for 1, 2, and 3 days were tested for thrips control. The 3-day ULO treatment achieved complete control of thrips. The treatment was also applied to Thompson seedless and Flame seedless grape varieties and grape quality was evaluated two weeks after the treatment. The ULO treatment had no negative impact on the visual quality of grapes. There were no significant differences in overall grape quality or premium quality berries between the ULO treatment and the control. The results indicated that the ULO treatment was effective and safe to control western flower thrips on table grapes and had a potential for commercial use.     

Stability of phycocyanin extracted from Spirulina sp.: Influence of temperature,pH and preservatives

Temperature and pH play an important role in the stability of phycocyanin, a natural blue colorant. Systematic investigations showed the maximum stability of phycocyanin was in the pH range 5.5–6.0. Incubation at temperatures between 47 and 64 °C caused the concentration (C_R) and half-life of phycocyanin in solution to decrease rapidly. The C_R value remained at approximately 50% after incubating for 30 min at 59 °C. After heating at 60 °C for 15 min, the C_R value of phycocyanin at pH 7.0 was maintained at around 62–70% when 20–40% glucose or sucrose was added, and the half-life increased from 19 min to 30–44 min. 2.5% sodium chloride was found to be an effective preservative for phycocyanin at pH 7.0 as a C_R value of 76% was maintained and the half-life of 67 min was increased.     

Expression of the 1-SST and 1-FFT genes and consequent fructan accumulation in Agave tequilana and A. inaequidens is differentially induced by diverse (a)biotic-stress related elicitors

The expression of genes coding for sucrose:sucrose 1-fructosyltransferase (1-SST; EC 2.4.1.99) and fructan:fructan 1-fructosyltransferase (1-FFT; EC 2.4.1.100), both fructan biosynthesizing enzymes, characterization by TLC and HPAEC-PAD, as well as the quantification of the fructo-oligosaccharides (FOS) accumulating in response to the exogenous application of sucrose, kinetin (cytokinin) or other plant hormones associated with (a)biotic stress responses were determined in two Agave species grown in vitro, domesticated Agave tequilana var. azul and wild A. inaequidens. It was found that elicitors such as salicylic acid (SA), and jasmonic acid methyl ester (MeJA) had the strongest effect on fructo-oligosaccharide (FOS) accumulation. The exogenous application of 1 mM SA induced a 36-fold accumulation of FOS of various degrees of polymerization (DP) in stems of A. tequilana. Other treatments, such as 50 mM abscisic acid (ABA), 8% Sucrose (Suc), and 1.0 mg L⁻¹ kinetin (KIN) also led to a significant accumulation of low and high DP FOS in this species. Conversely, treatment with 200 μM MeJA, which was toxic to A. tequilana, induced an 85-fold accumulation of FOS in the stems of A. inaequidens. Significant FOS accumulation in this species also occurred in response to treatments with 1 mM SA, 8% Suc, and 10% polyethylene glycol (PEG). Maximum yields of 13.6 and 8.9 mg FOS per g FW were obtained in stems of A. tequilana and A. inaequidens, respectively. FOS accumulation in the above treatments was tightly associated with increased expression levels of either the 1-FFT or the 1-SST gene in tissues of both Agave species.     

Effects of plant growth regulators and sucrose on post harvest physiology,membrane stability and vase life of cut spikes of gladiolus

Effects of post-harvest application of two plant growth regulators viz., gibberellic acid (GA₃) and benzyl adenine (BA) with sucrose in the vase solution on cell membrane stability and vase life of gladiolus were investigated. The vase solution treatment combinations of GA₃ and BA with sucrose significantly increased the membrane stability index and enhanced the vase life as compared to the sucrose alone treatments or the controls. Vase solution treatment of GA₃ (50 mg l⁻¹), followed by BA (50 mg l⁻¹) with sucrose (50 g l⁻¹) significantly increased solution uptake, fresh weight and dry weight of cut spikes. The same treatments also enhanced the concentration of reducing and non-reducing sugars in gladioli petals 4 days after treatment (DAT). Cut spikes in vase solution enriched with 50 mg l⁻¹ GA₃ + 50 g l⁻¹ sucrose showed higher antioxidative enzyme activities of superoxide dismutase (SOD) and glutathione reductase (GR), lower lipoxygenase (LOX) activity and lipid peroxidation (measured as TBARS). Petal membrane stability index was also highest in cut spikes 6 DAT with 50 mg l⁻¹ GA₃ + 50 g l⁻¹ sucrose vase solution. Treatment of gladiolus cut spikes with 50 mg l⁻¹ GA₃ + 50 g l⁻¹ sucrose vase solution showed two fold increase in vase life and improved flower quality with a higher number of open flower per spike at any one time. These results suggest that post-harvest application of GA₃ (50 mg l⁻¹) with sucrose (50 g l⁻¹) maintains higher spike fresh and dry weight, improves anti-oxidative defence, stabilizes membrane integrity leading to a delay in petal cell death.     

小苍兰切花不完全开放的原因探讨及防止措施

本文探明了小苍兰切花瓶插过程中小花不完全开放的原因,是由于采收时切花体内自身糖分积累不足所致。用高浓度(10%,20%,30%)蔗糖液处理,可增加切花各主、侧花序含糖量,提高开放率并延长瓶插寿命。同时还研究了几种含糖、化学药剂及激素的瓶插液对小花开放、花色和瓶插寿命的效应,以S+8-HQC+KT(蔗糖+8-羟基喹啉柠檬酸盐+激动素)效果较佳。此外,还比较了糖液处理和化学药剂保鲜两种方法的利弊和应用范围。     

Optimization of the cryopreservation of African clawed frog (Xenopus laevis) sperm

Cryopreservation of testicular sperm in the African clawed frog, Xenopus laevis, was tested using three penetrating cryoprotectants (DMSO, methanol, and glycerol) and three semen diluents (300 mmol/L glucose, 300 mmol/L sucrose, and a motility inhibiting saline [MIS] solution [150 mmol/L NaCl, 3 mmol/L KCL, 1 mmol/L Mg₂SO₄, 1 mmol/L CaCl₂, and 20 mmol/L Tris, pH 8.0]). Three freezing rates and four thawing rates were also tested, and the best freezing/thawing conditions have been determined. The responses of sperm motility, viability, and fertility were assessed. Incubation of the sperm macerates with penetrating cryoprotectants showed that DMSO was the least toxic and methanol the most toxic. Semen in cryodiluents frozen 10 cm above the surface of liquid nitrogen (freezing rate of 20 to 25 °C/min) and thawed at room temperature for 40 sec had significantly higher percentages of motile and viable sperm than that of semen frozen 5 cm or 8 cm above the surface of liquid nitrogen and thawed at 5, 25, or 30 °C for 10, 15, or 60 sec, respectively. Sperm frozen in MIS containing 5% DMSO had a higher hatching rate than that of sperm frozen in sucrose and glucose diluents containing 5% or 10% DMSO and in MIS containing 10% DMSO. Addition of 73 mmol/L sucrose to the sperm extender MIS + 5% DMSO could improve the postthaw sperm motility and fertility. In conclusion, dilution of collected sperm in MIS solution (to have a final concentration of 6.5 × 10⁶ to 8 × 10⁶/mL) containing 5% DMSO and 73 mmol/L sucrose, freezing in a vapor of liquid nitrogen at 10 cm above the surface, and thawing at room temperature for 40 sec was the best cryopreservation protocol. This protocol gave 70% hatching rate, 80% motility rate, and 75% viability rate of fresh hormonally induced sperm.     

Optimization of osmotic shock process variables for enhancement of the release of periplasmic interferon-α2b from Escherichia coli using response surface method

The osmotic shock process for the release of periplasmic recombinant human interferon-α2b from Escherichia coli was optimized using response surface method (RSM). The process parameters such as pH, buffer concentration and sucrose concentration in hypertonic solution, cell concentration to hypertonic solution, contact time of cells with hypertonic solution, temperature of hypertonic solution, cell concentration to hypotonic solution, contact time of cells with hypotonic solution and temperature of hypotonic solution were initially screened using Plackett Burman design. Further optimization was carried out using central composite design (one of the design in RSM) for sucrose concentration in hypertonic solution as well as cell concentration to hypertonic and hypotonic solutions. The optimal cell concentration was 0.05 g/mL in hypertonic solution and 0.2 g/mL in hypotonic solution. The use of hypertonic solution containing 18% sucrose with a combination of 100 mM Tris and 2.5 mM EDTA buffer (pH 8.0 and 25 °C) and cold water (4 °C) as a hypotonic solution gave the optimum release of interferon-α2b. Increased product concentration in the final solution resulted from the optimized process would reduce the downstream steps during purification. The concept of reuse of hypertonic solution was also demonstrated.     

The combined effects of gibberellic acid and salinity on some antioxidant enzyme activities,plant growth parameters and nutritional status in maize plants

The combined effects of salt stress and gibberellic acid (GA₃) on plant growth and nutritional status of maize (Zea mays L. cv., DK 647 F1) were studied in a pot experiment. Treatments were (1) control (C): nutrient solution alone, (2) salt stress (S): 100 mM NaCl, (3) S + GA1: 100 mM NaCl and 50 ppm GA₃ and (4) S + GA2: 100 mM NaCl and 100 ppm GA₃. Salt stress (S) was found to reduce the total dry matter, chlorophyll content, relative water content (RWC), but to increase proline accumulation, superoxide dismutase (SOD; EC 1.15.1.1), peroxidase (POD; EC 1.11.1.7) and polyphenol oxidase (PPO; 1.10.3.1) enzyme activities and electrolyte leakage. GA₃ treatments overcame to variable extents the adverse effects of NaCl stress on the above physiological parameters. GA₃ treatments reduced the activities of enzyme in the salt-stressed plants. Salt stress reduced some macro and micronutrient concentrations but exogenous application of GA₃ increased these to levels of control treatment. Foliar application of GA₃ counteracted some of the adverse effects of NaCl salinity with the accumulation of proline which maintained membrane permeability and increased macro and micronutrient levels.     

保鲜剂对夏季香石竹切花衰老的延缓作用

香石竹切花瓶插期间花瓣中ＳＯＤ和蛋白质在前期上升,然后比处理组提前３天呈下降趋势。采后花瓣中ＣＡＴ、可溶性总糖和蔗糖均为下降。经由蔗糖、８－羟基喹啉和植物生长调节剂或钴镍盐组成的保鲜剂能延缓切花花瓣中ＳＯＤ、ＣＡＴ活性的下降,阻碍蛋白质、可溶性总糖和蔗糖的降解速度,同时使切花的瓶插寿命延长、含水量增加。     

Development of a modified vitrification strategy suitable for subsequent scale-up for hepatocyte preservation

This work explores the design of a vitrification solution (VS) for scaled-up cryopreservation of hepatocytes, by adapting VS_basic (40% (v/v) ethylene glycol 0.6 M sucrose, i.e. 7.17 M ethylene glycol 0.6 M sucrose), previously proven effective in vitrifying bioengineered constructs and stem cells. The initial section of the scale-up study involved the selection of non-penetrating additives to supplement VS_basic and increase the solution’s total solute concentration. This involved a systematic approach with a step-by-step elimination of non-penetrating cryoprotectants, based on their effect on cells after long/short term exposures to high/low concentrations of the additives alone or in combinations, on the attachment ability of hepatocytes after exposure. At a second stage, hepatocyte suspension was vitrified and functions were assessed after continuous culture up to 5 days.Results indicated Ficoll as the least toxic additive. Within 60 min, the exposure of hepatocytes to a solution composed of 9% Ficoll + 0.6 M sucrose (10⁻³ M Ficoll + 0.6 M sucrose) sustained attachment efficiency of 95%, similar to control. Furthermore, this additive did not cause any detriment to the attachment of these cells when supplementing the base vitrification solution VS_basic. The addition of 9% Ficoll, raised the total solute concentration to 74.06% (w/v) with a negligible 10⁻³ M increase in molarity of the solution. This suggests main factor in inducing detriment to cells was the molar contribution of the additive.Vitrification protocol for scale-up condition sustained hepatocyte suspension attachment efficiency and albumin production. We conclude that although established approach will permit scaling-up of vitrification of hepatocyte suspension, vitrification of hepatocytes which are attached prior to vitrification is more effective by comparison.