Variation of the HES Concentration for the Cryopreservation of Keratinocytes in Suspensions and in Monolayers

Pasch, J.; Schiefer, Ana Iris; Heschel, I.; Dimoudis, Nikolaos; Rau, G.

doi:10.1006/cryo.2000.2270

Cited by 29 publications

(17 citation statements)

References 4 publications

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“…It was previously shown by Pasch et al that HES can be used to optimally cryopreserve human keratinocytes in suspension and in monolayers [20, 21]. Our data for HaCaT cells corroborate these findings, showing that both cell numbers (Fig.…”

Section: Discussionsupporting

confidence: 90%

“…HES has been previously used to cryopreserve keratinocytes [20, 21], islets [22], red blood cells [23, 24], peripheral blood stem cells [25, 26], and other cell types [27, 28]. The addition of HES to the CPA solution has been reported to increase the recovery and viability rate after freezing [21]. In our recent work, we successfully cryopreserved rat mesenchymal stem cells using a 5% DMSO, 5% HES solution [29].…”

Section: Introductionmentioning

confidence: 99%

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Cryopreservation of dermal fibroblasts and keratinocytes in hydroxyethyl starch–based cryoprotectants

et al. 2016

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BackgroundPreservation of human skin fibroblasts and keratinocytes is essential for the creation of skin tissue banks. For successful cryopreservation of cells, selection of an appropriate cryoprotectant agent (CPA) is imperative. The aim of this study was to identify CPAs that minimize toxic effects and allow for the preservation of human fibroblasts and keratinocytes in suspension and in monolayers.ResultsWe cryopreserved human fibroblasts and keratinocytes with different CPAs and compared them to fresh, unfrozen cells. Cells were frozen in the presence and absence of hydroxyethyl starch (HES) or dimethyl sulfoxide (DMSO), the latter of which is a commonly used CPA known to exert toxic effects on cells. Cell numbers were counted immediately post-thaw as well as three days after thawing. Cellular structures were analyzed and counted by labeling nuclei, mitochondria, and actin filaments. We found that successful cryopreservation of suspended or adherent keratinocytes can be accomplished with a 10% HES or a 5% HES, 5% DMSO solution. Cell viability of fibroblasts cryopreserved in suspension was maintained with 10% HES or 5% HES, 5% DMSO solutions. Adherent, cryopreserved fibroblasts were successfully maintained with a 5% HES, 5% DMSO solution.ConclusionWe conclude that skin tissue cells can be effectively cryopreserved by substituting all or a portion of DMSO with HES. Given that DMSO is the most commonly used CPA and is believed to be more toxic than HES, these findings are of clinical significance for tissue-based replacement therapies. Therapies that require the use of keratinocyte and fibroblast cells, such as those aimed at treating skin wounds or skin burns, may be optimized by substituting a portion or all of DMSO with HES during cryopreservation protocols.

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Section: Discussionsupporting

confidence: 90%

Section: Introductionmentioning

confidence: 99%

Cryopreservation of dermal fibroblasts and keratinocytes in hydroxyethyl starch–based cryoprotectants

et al. 2016

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“…Membrane-permeating CPAs such as glycerol can prevent intracellular ice formation, but repeated washing steps are necessary to remove it before fat grafting [9]. HES is a nonmembrane-permeating CPA [15,16], so it is less toxic to cells and therefore is an ideal agent for clinical use. Wolter et al [9] compared DMEM plus 10% HES, DMEM plus 10% glycerol, and DMEM plus 10% glycerol, 5% dextran, 5% polyvinyl pyridine (PVP) and found that the membrane-permeating protectants seem to be beneficial.…”

Section: Discussionmentioning

confidence: 99%

Cryopreservation of Fat Tissue and Application in Autologous Fat Graft: In Vitro and In Vivo Study

Liao

et al. 2011

Aesth Plast Surg

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“…Keratinocyte is a fastidious cell type and easily loses its growth potential in vitro 13. Although the long‐term cryopreservation of cultured keratinocytes for later application was reported previously,14 but the manipulation processes of cell cryopreservation and refreshing are often complex and would result some cell loss. It has been described that the skin had been stored in 4°C for a short period and then refreshed for the cultivation of keratinocytes 15.…”

Section: Introductionmentioning

confidence: 99%

Cryopreserved lip mucosa tissue derived keratinocytes can fabricate tissue engineered palatal mucosa equivalent

Xiong

Jia

et al. 2010

J Biomed Mater Res

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Clinical application of tissue engineered palatal mucosa is hampered by unavailability of suitable oral keratinocytes as seeding cells. The aim of this study is to fabricate a tissue engineered palatal mucosa equivalent from the oral keratinocytes which cultured from cryopreserved lip mucosa tissues. Abundant lip mucosa tissues during cheilorrhaphy were firstly cryopreserved in liquid nitrogen for four to six months, and then recovered to culture oral keratinocytes for the fabrication of oral mucosa equivalent. In the control groups, oral keratinocytes cultured from fresh lip mucosa, fresh palate mucosa, and cryopreserved palate mucosa were used to fabricate oral mucosa equivalents. Attachment rate of the oral keratinocytes derived from cryopreserved lip mucosa was lower than that of the keratinocytes from fresh lip mucosa samples, however, the cell cycle distribution of oral keratinocytes cultured from all four groups of mucosa samples were similar. Histologically, the fabricated mucosa equivalents from these four groups had four- to six epithelial layers, the basal cells were cubic and the outmost cells were flatten with narrow nuclei which paralleled to the surface of the dermal matrix. Additionally, Ki-67 positive stained cells were mainly located in the basal layer of the epithelium of these equivalents. These characteristics disclosed that the oral mucosa equivalent cultured from the cryopreserved lip mucosa tissue was not different with the equivalents from other groups and similar to the native palate mucosa tissue. It suggested that the cryopreserved lip mucosa tissues could be used for the construction of palatal mucosal equivalent for clinical application.

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Variation of the HES Concentration for the Cryopreservation of Keratinocytes in Suspensions and in Monolayers

Cited by 29 publications

References 4 publications

Cryopreservation of dermal fibroblasts and keratinocytes in hydroxyethyl starch–based cryoprotectants

Cryopreservation of dermal fibroblasts and keratinocytes in hydroxyethyl starch–based cryoprotectants

Cryopreservation of Fat Tissue and Application in Autologous Fat Graft: In Vitro and In Vivo Study

Cryopreserved lip mucosa tissue derived keratinocytes can fabricate tissue engineered palatal mucosa equivalent

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