Use of different cooling rates during freezing to separate populations of human peripheral blood lymphocytes

Farrant, J.; Knight, Stella C.; Morris, G.J.

doi:10.1016/0011-2240(72)90173-3

Cited by 51 publications

(7 citation statements)

References 36 publications

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“…The " slow-freezing " and " fast-freezing " rates used by Levy were not defined but were probably intermediate between the slow-freezing and fast-freezing rates used in this study. Levy used 10% glycerol as the cryoprotective agent, whereas we used 7 3 % DMSO; preservation of viability with DMSO is usually favoured by slow freezing rates (Thurston and Harris, 1970;Farrant, Knight and Morris, 1972;Whittingham, Leibo and Mazur, 1972 ;Beadle and Harris, 1974).…”

Section: The Effect Of Freezing Ratementioning

confidence: 99%

The Effect of Freezing and Storage in Liquid Nitrogen on the Viability and Growth of Mycobacterium Leprae

Colston¹,

Hilson

1979

Journal of Medical Microbiology

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THE use of liquid nitrogen as a means of preserving biological material has recently been applied to suspensions of Mycobacterium leprae (Pattyn, 1973). A satisfactory method of preserving M . leprae in vitro would permit the use of standardised inocula for a series of experiments in vivo instead of individual inocula from mouse-passage material. However, previous attempts to preserve these organisms at low temperatures have resulted in serious reduction or total loss of viability (Levy, 1969 and1971). Pattyn reported that, by freezing M . leprae suspended in a medium containing 7.5% dimethyl sulphoxide (DMSO) in liquid nitrogen, infectivity of the suspensions when inoculated into the footpads of mice was retained, but no quantitative determination of the effect of such treatment on the viability and subsequent growth of M. Ieprae was made. This paper reports observations on the effects of freezing rate, prolonged storage in liquid nitrogen, and thawing rate on the viability and subsequent growth of M. leprae. MATERIALS AND METHODS Strains of M . leprae andpreparation of inoculaThree strains of M . leprae were used: SBL16220, SBL15337, and TG, all originally derived from untreated lepromatous leprosy patients and subsequently maintained in mouse passage in our laboratory. The inoculation of female ASH/CSI mice (Charles River UK Ltd), harvesting and counting of M . leprae from mouse footpads and assessment of growth were performed according to previously reported methods (Holmes and Hilson, 1972).Inocula were prepared and diluted in Bacto TB Broth (Difco) with 7.5 % DMSO (v/v).

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Section: The Effect Of Freezing Ratementioning

confidence: 99%

The Effect of Freezing and Storage in Liquid Nitrogen on the Viability and Growth of Mycobacterium Leprae

Colston¹,

Hilson

1979

Journal of Medical Microbiology

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“…Pegg (1965) used somewhat slower cooling to preserve human lymphocytes in 1-4 M DMSO. The interaction between DMSO concentration and cooling rate was studied by Farrant et al (1972), who found that the maximum recovery of CON-A responsive human lymphocytes was obtained at 0-3°C/min in 1-4 M DMSO and at 1°C min-' in 0-7 M DMSO. Thorpe et al (1975) have studied the influence of post-thaw manipulations on the recovery of frozen mouse lymphocytes: they found that dilution of the suspension to remove DMSO should be carried out at 25°C rather than at 0°C, and that the presence of 10 % serum, and the use of low g forces for centrifugation, were beneficial.…”

Section: Preservation Of Lymphocytesmentioning

confidence: 99%

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

Pegg¹

1976

J Clin Pathol

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“…Typically, this will involve the addition of cryoprotectants, most commonly dimethyl sulphoxide (DMSO), dextran, and hydroxyethyl starch, to protect cells during the cryopreservation process [7À10]. Samples are cooled in a controlled-rate freezer (CRF) following a specific cooling profile, because the importance of a precisely controlled cooling rate in mammalian cell cryopreservation has long been recognized [11,12].…”

Section: Introductionmentioning

confidence: 99%

“…Excessive supercooling is known to be harmful to many cell types [16] and can give results for a QC sample that do not reflect the likely performance of the comparable bulk sample after thawing [9,10,13À16]. Cord blood samples contain a heterogeneous population of cells, and because different cell types can respond differently to ice nucleation, a difference in post-thaw cell populations between recovered QC and bulk samples may also occur [9,11,16,17]. Although such QC segments are commonly used in cord blood cryopreservation, their use in other procedures must be considered as the use of cryopreserved products in cell therapies increases [15,18,19].…”

Section: Introductionmentioning

confidence: 99%

Recovery and Post-Thaw Assessment of Human Umbilical Cord Blood Cryopreserved as Quality Control Segments and Bulk Samples

Kilbride

Meneghel

Lamb

et al. 2019

Biology of Blood and Marrow Transplantation

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Quality control (QC) segments conjoined to a bulk sample container are used to evaluate the viability and quality of cryopreserved umbilical cord blood (UCB). Such QC segments are typically attached lengths of sealed tubing that are cooled concurrently with the bulk sample, both containing material from the same donor. QC segments are thawed independently of the bulk sample to assess the quality of the cryopreserved product. In current practice, there is typically post-thaw variation between the QC segment and the bulk sample which if suggestive of inadequate performance, could lead to material being needlessly discarded. In this study, these performance differences were quantified. Two cooling protocols in common use, 1 with and 1 without a "plunge" step to induce ice nucleation, gave equivalent results that maintained the QC segment versus bulk sample differences. Ice nucleated at significantly lower temperatures in the QC segments compared with the bulk samples, a consequence of their lower volume, thereby enhancing damaging osmotic stress. A reduction in total viable cells of approximately 10% was recorded in the QC segments compared with comparable bulk samples. It has been shown that CD45 + cells are more adversely impacted by this lower ice nucleation temperature than CD34 + cells, which can result in altered composition of the post-thaw cell population.

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Use of different cooling rates during freezing to separate populations of human peripheral blood lymphocytes

Cited by 51 publications

References 36 publications

The Effect of Freezing and Storage in Liquid Nitrogen on the Viability and Growth of Mycobacterium Leprae

The Effect of Freezing and Storage in Liquid Nitrogen on the Viability and Growth of Mycobacterium Leprae

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

Recovery and Post-Thaw Assessment of Human Umbilical Cord Blood Cryopreserved as Quality Control Segments and Bulk Samples

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