The origin, ultrastructure, and microbiology of the sediment accumulating in liquid nitrogen storage vessels

Morris, G.J.

doi:10.1016/j.cryobiol.2005.01.005

Cited by 83 publications

(74 citation statements)

References 8 publications

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“…The possibility of cross-contamination may be due to possible sources for the presence of organisms in LN: their entrance from external environment or their derivation from the common source of LN (Morris 2005;Bielanski 2005). LN is manufactured by the condensation of atmospheric nitrogen under high pressure.…”

Section: Discussionmentioning

confidence: 99%

Microbiological analysis of cryopreserved human heart valves after storage: a survey of 3 banks in Spain

Villalba¹,

Mirabet²,

Rendal

et al. 2009

Cell Tissue Bank

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Several reports have shown liquid nitrogen containers as not being sterile. Microorganism transmission has been observed in different cells and tissues stored under this condition, but there is no data on contamination of stored human valves. We performed a survey on heart valve banking in Spain. Regarding the questionnaire, we have a complete microbiological analysis of 304 thawed tissues prior to implant. In six cases positive culture results were observed. Patient follow-up did not reveal any adverse effects. Although some other possibilities should be stated, contamination of heart valves during storage in liquid nitrogen should be considered as a risk element in tissue banking. Strategies to asses and prevent microbial transmission from liquid nitrogen to heart valve banking ought to be further developed.

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Section: Discussionmentioning

confidence: 99%

Microbiological analysis of cryopreserved human heart valves after storage: a survey of 3 banks in Spain

Villalba¹,

Mirabet²,

Rendal

et al. 2009

Cell Tissue Bank

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“…2 Cryopreservation in bags of large volumes ( > 100 mL) of adult stem cells 3 and of mammalian tissue culture cells [4][5][6] using conventional liquid nitrogen-based controlled rate freezers (CRFs) is established. However, the use of liquid nitrogen is problematic in a GMP environment; following manufacture, liquid nitrogen contains very low levels of contaminants, but during transport and storage it can become contaminated by ice, inanimate debris, and viable microorganisms [7][8][9] that can transfer to the local environment. 7,10 It is possible to filter industrial quantities of liquid nitrogen to appropriate standards and the UV sterilization of liquid nitrogen has been proposed 11 ; however, unless liquid nitrogen is filtered or sterilized by validated methods it cannot be used in a clean room without compromising air quality.…”

Section: Introductionmentioning

confidence: 99%

GMP Cryopreservation of Large Volumes of Cells for Regenerative Medicine: Active Control of the Freezing Process

MassieIsobel¹,

Selden²,

HodgsonHumphrey³

et al. 2014

Tissue Engineering Part C: Methods

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Cryopreservation protocols are increasingly required in regenerative medicine applications but must deliver functional products at clinical scale and comply with Good Manufacturing Process (GMP). While GMP cryopreservation is achievable on a small scale using a Stirling cryocooler-based controlled rate freezer (CRF) (EF600), successful large-scale GMP cryopreservation is more challenging due to heat transfer issues and control of ice nucleation, both complex events that impact success. We have developed a large-scale cryocoolerbased CRF (VIA Freeze) that can process larger volumes and have evaluated it using alginate-encapsulated liver cell (HepG2) spheroids (ELS). It is anticipated that ELS will comprise the cellular component of a bioartificial liver and will be required in volumes of *2 L for clinical use. Sample temperatures and Stirling cryocooler power consumption was recorded throughout cooling runs for both small (500 mL) and large (200 mL) volume samples. ELS recoveries were assessed using viability (FDA/PI staining with image analysis), cell number (nuclei count), and function (protein secretion), along with cryoscanning electron microscopy and freeze substitution techniques to identify possible injury mechanisms. Slow cooling profiles were successfully applied to samples in both the EF600 and the VIA Freeze, and a number of cooling and warming profiles were evaluated. An optimized cooling protocol with a nonlinear cooling profile from ice nucleation to -60°C was implemented in both the EF600 and VIA Freeze. In the VIA Freeze the nucleation of ice is detected by the control software, allowing both noninvasive detection of the nucleation event for quality control purposes and the potential to modify the cooling profile following ice nucleation in an active manner. When processing 200 mL of ELS in the VIA Freeze-viabilities at 93.4% -7.4%, viable cell numbers at 14.3 -1.7 million nuclei/mL alginate, and protein secretion at 10.5 -1.7 mg/ mL/24 h were obtained which, compared well with control ELS (viability -98.1% -0.9%; viable cell numbers -18.3 -1.0 million nuclei/mL alginate; and protein secretion -18.7 -1.8 mg/mL/24 h). Large volume GMP cryopreservation of ELS is possible with good functional recovery using the VIA Freeze and may also be applied to other regenerative medicine applications.

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“…During storage over a long period of time, ice sediment may accumulate inside the tank. This sediment can be a source of contamination for cryopreserved biological tissues (Morris, 2005). Thibier and Guerin (2000) described that such contaminants can be present in extenders, on equipment, and in the liquid nitrogen as well.…”

Section: Discussionmentioning

confidence: 99%

“…It is known that contamination can occur during tissue, semen or embryo storage in liquid nitrogen and the cryopreservation of external microorganisms can occur by extravasation of cryoprotectors used in semen from damaged straws (Piaseka-Serafin, 1972;Bielansky et al, 2003;Bielanski, 2005a;Morris, 2005). As water vaporizes, cools and freezes above an open liquid-nitrogen tank, small ice crystals with a high electrostatic charge are formed and capture airborne microorganisms which fall into the tank (Grout and Morris, 2009).…”

Section: Introductionmentioning

confidence: 99%

Decontamination of naturally contaminated liquid nitrogen storage tanks

et al. 2014

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-The objective of this study was to evaluate the efficacy of cleaning and decontamination procedures in liquid nitrogen tanks. We evaluated 151 canisters and 133 bottoms from 133 nitrogen tanks of companies or farms for the presence of bacteria and fungi. Samples were collected from the canisters and the bottom of tanks containing liquid nitrogen. Tanks were divided into Group 1 (G1): tanks decontaminated with 2% glutaraldehyde -Glutaron ® II (n = 16 canisters in 8 tanks); Group 2 (G2): decontamination with 70% ethanol (n = 20 canisters in 10 tanks); and Group 3 (G3): decontamination with 70% ethanol (n = 115 canisters in 115 tanks). Tanks in Groups 1 and 2 belonged to companies; Group 3 tanks belonged to farms. The culture of canisters showed twelve genera of bacteria and five genera of fungi. Bacillus cereus was the most prevalent bacterial contaminant (42/133) in liquid nitrogen tanks (31.57%). Decontamination by 2% glutaraldehyde plus 70% ethanol was effective and no difference was found between the decontamination methods of Groups 1 and 2. In Group 3 the decontamination method was considered effective. Handling procedures with high hygienic standards should be recommended to avoid contamination of liquid nitrogen tanks on farms.

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The origin, ultrastructure, and microbiology of the sediment accumulating in liquid nitrogen storage vessels

Cited by 83 publications

References 8 publications

Microbiological analysis of cryopreserved human heart valves after storage: a survey of 3 banks in Spain

Microbiological analysis of cryopreserved human heart valves after storage: a survey of 3 banks in Spain

GMP Cryopreservation of Large Volumes of Cells for Regenerative Medicine: Active Control of the Freezing Process

Decontamination of naturally contaminated liquid nitrogen storage tanks

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