Long-term preservation of cells and tissues: a review.

Pegg, D.E.

doi:10.1136/jcp.29.4.271

Cited by 65 publications

(28 citation statements)

References 81 publications

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“…It is based upon the formation of intracellular ice crystals, which can be demonstrated by light and electron microscopy (29). Usually, two or three (3,4), some·t imes four cycles (20) of freezing and thawing are described, sometimes followed by an additional homogenization in the Potter-Ehehjem (11) homogenizer.…”

Section: Homogenizationmentioning

confidence: 99%

The Measurement of Enzyme Activities in the Resting Human Polymorphonuclear Leukocyte — Critical Estimate of a Method

Fauth

Schlechtriemen²,

Heinrichs³

et al. 1993

Clinical Chemistry and Laboratory Medicine

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As a system for study, the isolated human polymorphonuclear leukocyte combines the advantages of a quasi-non-invasive preparation with a nearly complete complement of enzymes of carbohydrate and energy metabolism. However, small sample volumes and, in some cases, very low enzyme activities make high demands on sample processing, storage, and performance of continuous measurements, if the enzyme activities are to be measured with acceptable reproducibility. In the presented study several aspects of homogenization, storage, and continuous measurement were scrutinized, to identify critical steps and consider ways of optimizing the method. Polymorphonuclear leukocytes were separated from the blood of healthy subjects by sedimentation and density gradient centrifugation. After ultrasonic homogenization, 13 enzymes of glycolysis and gluconeogenesis, the tricarboxylic acid cycle, and glycogen metabolism were determined photometrically. The variation of several conditions showed: 1. The duration of exposure to ultrasound for the homogenization of polymorphonuclear leukocytes has no influence over a wide range of time.2. Addition of the detergents Triton X-100 and deoxycholic acid, as well as the SH-group protector dithiothreitol, to the homogenizing medium increased the measured activities of only a few enzymes.3. Considerable inaccuracy was encountered-when the suspension was divided into parts for homogenization with different additives; such splitting of the suspension should therefore be performed only when necessary, as in the determination of reference values (e. g. protein or DNA content of the cell suspension).4. Twenty four-fold determination of enzyme activities from one homogenate resulted in precisions between 4.5% (citrate synthase) and 14.4% (transketolase), which is satisfactory for the low activities (as low as 1 U/l) in the homogenate. 5. The reproducibility of enzyme activities, measured in homogenates of polymorphonuclear leukocytes from different blood samples drawn simultaneously, was only slightly worse than that of the continuous measurement method itself. Thus, the precision of the measurement of enzyme activity seems to be the main determinant of the overall method. In conclusion, the described procedure of separation, homogenization, and enzyme measurement in human polymorphonuclear leukocyte meets the requirements of biochemical or clinical trials and can be recommended for clinical metabolic studies.

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Section: Homogenizationmentioning

confidence: 99%

The Measurement of Enzyme Activities in the Resting Human Polymorphonuclear Leukocyte — Critical Estimate of a Method

Fauth

Schlechtriemen²,

Heinrichs³

et al. 1993

Clinical Chemistry and Laboratory Medicine

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“…So far, there has been only little informa tion on the behaviour of enzymes of human serum at lower temperature than -20 °C [10,14], The good experience gained in preserva tion of cells in liquid nitrogen [11] led us to study the long-term stability of some en zymes of human serum under these storage…”

mentioning

confidence: 99%

Long-Term Stability of Enzymes in Human Serum Stored in Liquid Nitrogen

Jung¹,

Bader²,

Grützmann³

1984

Enzyme

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We analyzed the stability of the enzymes α-amylase (EC 3.2.1.1), alkaline phosphatase (EC 3.1.3.1), alanine aminotransferase (EC 2.6.1.2), aspartate aminotransferase (EC 2.6.1.1), creatine kinase (EC 2.7.3.2), glutamate dehydrogenase (EC 1.4.1.3), γ-glutamyltransferase (EC 2.3.2.2) and lactate dehydrogenase (EC 1.1.1.27) of a human serum pool during storage in liquid nitrogen for a period of 10 months. Except amylase and creatine kinase, all enzymes were stable. Amylase increased in activity, creatine kinase activity decreased. Therefore, human serum stored at -196 °C can be used as satisfactory substitute for lyophilized enzyme control serum in internal quality control and stable enzyme material for optimization of methods.

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“…The first evidence that animals could be protected from marrow lethal therapy by the use of cryopreserved bone marrow was generated in 1954 (19), but the physiology of cryopreservation is still not fully understood. There are many recent reviews which explore cryopreservation and its application to the storing of bone marrow (20)(21)(22)(23). After harvest, marrow is centrifuged to separate nucleated cells from erythrocytes.…”

Section: Autologous Bone Marrow Infusion As Support: Technical Aspectsmentioning

confidence: 99%

High dose chemotherapy with autologous marrow rescue in the treatment of resistant solid tumors

LeMaistre

Knight

1983

Invest New Drugs

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Toxicity in the form of marrow suppression has hampered the investigation of intensive chemotherapy as it applies to the treatment of solid tumors. The use of autologous marrow rescue to ameliorate protracted aplasia permits the application of high dose chemotherapy to the treatment of solid tumors. We review the theoretical limitations of this technique and present results reported in selected solid tumors. Autologous marrow rescue from intensive chemotherapy is an innovative approach in overcoming the resistence to chemotherapy that is typical of solid tumors. The role of this therapy in the treatment of solid tumors remains to be defined.

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Long-term preservation of cells and tissues: a review.

Abstract: There has been a steady growth of interest in cryobiology during the past 10 years: this decade has seen a considerable increase in our

Cited by 65 publications

References 81 publications

The Measurement of Enzyme Activities in the Resting Human Polymorphonuclear Leukocyte — Critical Estimate of a Method

The Measurement of Enzyme Activities in the Resting Human Polymorphonuclear Leukocyte — Critical Estimate of a Method

Long-Term Stability of Enzymes in Human Serum Stored in Liquid Nitrogen

High dose chemotherapy with autologous marrow rescue in the treatment of resistant solid tumors

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