Sucrose promotes the functional activity of blood vessels after cryopreservation in DMSO-containing fetal calf serum

Müller‐Schweinitzer, Else; Ellis, P.

doi:10.1007/bf00168954

Cited by 17 publications

(12 citation statements)

References 12 publications

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“…Lactobionate is a disaccharide derivative, and other disaccharides are known to act as cryoprotective agents in various cell types 53 ; the presence of the disaccharide sucrose in combination with DMSO has also been shown to improve functionality of cryopreserved vessels. 54 Lactobionate might stabilize the plasma membrane during cryopreservation as known for other disaccharides and/or the low concentration of chloride might avoid chloride influx via a destabilized plasma membrane/fluid-to-gel phase transition during freezing/thawing. 55 Chloride fluxes in the frozen state, in contrast, are unlikely.…”

Section: Discussionmentioning

confidence: 99%

Serum- and albumin-free cryopreservation of endothelial monolayers with a new solution

2018

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Cryopreservation is the only long-term storage option for the storage of vessels and vascular constructs. However, endothelial barrier function is almost completely lost after cryopreservation in most established cryopreservation solutions. We here aimed to improve endothelial function after cryopreservation using the 2D-model of porcine aortic endothelial cell monolayers. The monolayers were cryopreserved in cell culture medium or cold storage solutions based on the 4°C vascular preservation solution TiProtec®, all supplemented with 10% DMSO, using different temperature gradients. After short-term storage at −80°C, monolayers were rapidly thawed and re-cultured in cell culture medium. Thawing after cryopreservation in cell culture medium caused both immediate and delayed cell death, resulting in 11 ± 5% living cells after 24 h of re-culture. After cryopreservation in TiProtec and chloride-poor modifications thereof, the proportion of adherent viable cells was markedly increased compared to cryopreservation in cell culture medium (TiProtec: 38 ± 11%, modified TiProtec solutions ≥ 50%). Using these solutions, cells cryopreserved in a sub-confluent state were able to proliferate during re-culture. Mitochondrial fragmentation was observed in all solutions, but was partially reversible after cryopreservation in TiProtec and almost completely reversible in modified solutions within 3 h of re-culture. The superior protection of TiProtec and its modifications was apparent at all temperature gradients; however, best results were achieved with a cooling rate of −1°C/min. In conclusion, the use of TiProtec or modifications thereof as base solution for cryopreservation greatly improved cryopreservation results for endothelial monolayers in terms of survival and of monolayer and mitochondrial integrity.

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Section: Discussionmentioning

confidence: 99%

Serum- and albumin-free cryopreservation of endothelial monolayers with a new solution

2018

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“…The arteries were cleaned of loose connective tissue and used in organ bath studies either unfrozen or following cryostorage. For cryopreservation the arterial segments (10–20 mm long) were suspended in 2 ml liquid nitrogen storage ampules (Life Technologies AG, Basel, Switzerland) filled with 1.6 ml RPMI 1640 culture medium containing 1.8 M DMSO and 0.1 M sucrose ( Müler‐Schweinitzer & Ellis, 1992 ). After an equilibration time of 10–20 min at room temperature the samples were placed into a polystyrene box (wall thickness ∼ 10 mm) and slowly frozen in a freezer maintained at −70°C.…”

Section: Methodsmentioning

confidence: 99%

The mechanism of cryoinjury: In vitro studies on human internal mammary arteries

Müller‐Schweinitzer

Brett

Zerkowski

et al. 2000

British J Pharmacology

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1 The mechanism of cryoinjury was investigated in human internal mammary arteries (IMA) by monitoring contractile responses to ET-1 and KCl. 2 For cryopreservation segments of IMA were equilibrated for 20 min with the cryomedium (RPMI 1640 culture medium containing 1.8 M DMSO and 0.1 M sucrose), frozen at a mean cooling rate of 1.38C min 71 to 7708C and stored in liquid nitrogen. Before use, samples were thawed slowly and the cryomedium removed by dilution. 3 Compared to unfrozen controls, ET-1 stimulated frozen/thawed IMA with similar e cacy but at 3 fold lower concentrations (P50.001). Addition of ET-1 (100 nM) induced maximal contraction of unfrozen IMA within 10 min, declining thereafter to 25% after 90 min. In frozen/thawed IMA the ET-1-induced contraction was sustained but could be reversed if protein kinase C was blocked by staurosporine (100 nM). Responses to ET-1 of cryostored IMA were 5 fold more susceptible to blockade by nifedipine than those of controls. 4 After cryostorage the e cacy of KCl was diminished to 40% (P50.05) and the KCl curve was shifted to the left (2 fold, P50.001). In both unfrozen and cryostored IMA the KCl (60 mM) e ect was sustained and equally susceptible to nifedipine. 5 It is suggested that the smooth muscle cell of IMA is receptive to physical forces which occur during cryopreservation. These forces modify transmembrane signal transduction and intracellular pathways, that are common to pharmacological agonists thereby changing vascular responses to several contractile agonists after thawing. British Journal of Pharmacology (2000) 130, 636 ± 640 Keywords: Human internal mammary artery; cryopreservation; cryoinjury; endothelin-1; depolarization; protein kinase C activation; calcium in¯ux; staurosporine; nifedipine Abbreviations: ANOVA, one-way analysis of variance; DAG, diacylglycerol; DMSO, dimethylsulphoxide; EC 50 , concentration of an agonist producing 50% of E max ; EDTA, ethylenediaminetetraacetatic acid disodium salt; E max , maximal contractile e ect; ET-1, endothelin-1; IC 50 , drug concentration of an inhibitor producing 50% inhibition of E max of an agonist; IMA, internal mammary artery; IP 3 , phosphoinositide 4,5-trisphosphate; KH, Krebs-Henseleit; pD 2 , negative logarithm of the molar concentration of agonist producing 50% of E max ; PGF 2a , prostaglandin

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“…During microsurgery, papaverine is used as a smooth muscle relaxant and is directly applied to blood vessels [28,29]. It is also applied in cryopreservation of blood vessels along with other glycosaminoglycans and protein suspensions [21,30]. Papaverine also functions as a vasodilator during cryopreservation when used in conjunction with verapamil, phentolamine, nifedipine, tolazoline or nitroprusside [22,31].…”

Section: Economic Importance Of Opium Poppy and Its Derivativesmentioning

confidence: 99%

Opium Poppy: Genetic Upgradation Through Intervention of Plant Breeding Techniques

Mishra¹,

Rastogi²,

Siddiqui³

et al. 2013

Plant Breeding From Laboratories to Fields

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Sucrose promotes the functional activity of blood vessels after cryopreservation in DMSO-containing fetal calf serum

Cited by 17 publications

References 12 publications

Serum- and albumin-free cryopreservation of endothelial monolayers with a new solution

Serum- and albumin-free cryopreservation of endothelial monolayers with a new solution

The mechanism of cryoinjury: In vitro studies on human internal mammary arteries

Opium Poppy: Genetic Upgradation Through Intervention of Plant Breeding Techniques

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