Skin Banking: A Simple Method for Cryopreservation of Split-Thickness Skin and Cultured Human Epidermal Keratinocytes

Konstantinow, Alexander; Mühlbauer, W.; Hartinger, A.; Donnersmarck, G. G. H. Von

doi:10.1097/00000637-199101000-00014

Cited by 16 publications

(7 citation statements)

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“…In a small, self‐conducted, prospective clinical trial with eight patients, half of each DSW area (0·3 mm thickness) was covered with Biobrane® (Smith and Nephew, London, UK) and the other half with allogeneic cryopreserved keratinocytes. In the Biobrane‐covered areas, complete epithelialisation was observed after a mean of 13·4 days and in the wounds covered with CEA after a mean of 7·5 days (Table ) . CEA were also used by Phillips et al in the treatment of superficial DSWs in nine patients .…”

Section: Discussionmentioning

confidence: 99%

Effectiveness of collagen/oxidised regenerated cellulose/silver‐containing composite wound dressing for the treatment of medium‐depth split‐thickness skin graft donor site wounds in multi‐morbid patients: a prospective, non‐comparative, single‐centre study

Konstantinow¹,

Fischer²,

Ring³

2016

International Wound Journal

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Split-thickness skin grafting (STSG) is a widely used method in reconstructive surgery, but donor site wounds (DSWs) are often slow healing and painful. This prospective study evaluated the performance of a composite wound dressing containing collagen/oxidised regenerated cellulose in the treatment of medium-depth (0·4 mm) DSWs in 25 multi-morbid patients with chronic leg ulcers requiring STSG. The range of patients' ages was 44-84 years (mean 71·6 years) with DSW sizes ranging between 12 and 162 cm (mean 78 cm ). Comorbidities included anticoagulation therapy (15 patients), anaemia (11 patients), diabetes (6 patients) and methicillin-resistant Staphylococcus aureus (MRSA) ulcer colonisation (6 patients). The first dressing change was performed after 10 days. Complete reepithelialisation was observed between the 10th and 34th day (mean 17·2, median 14 days). Postoperative medium to strong bleeding occurred in only five patients (four with anticoagulation). Wound pain levels one day after harvesting were only moderate (range 0-1·5, mean 0·5, median 0·5 on a six-item scale). No wound infection was observed during the first dressing. The composite dressing used allowed for the fast healing of medium-depth DSWs with minimal or no postoperative pain and bleeding in older multi-morbid patients under anticoagulation treatment.

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Section: Discussionmentioning

confidence: 99%

Effectiveness of collagen/oxidised regenerated cellulose/silver‐containing composite wound dressing for the treatment of medium‐depth split‐thickness skin graft donor site wounds in multi‐morbid patients: a prospective, non‐comparative, single‐centre study

Konstantinow¹,

Fischer²,

Ring³

2016

International Wound Journal

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“…Glucose and lactate values in the storage media gave an indication of metabolic status and provided further evaluation of cell viability [26], [27], whereas oxygen tension and pH measurements ensured storage conditions were within a physiologically relevant range.…”

Section: Discussionmentioning

confidence: 99%

Effect of Storage Temperature on Cultured Epidermal Cell Sheets Stored in Xenobiotic-Free Medium

et al. 2014

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Cultured epidermal cell sheets (CECS) are used in regenerative medicine in patients with burns, and have potential to treat limbal stem cell deficiency (LSCD), as demonstrated in animal models. Despite widespread use, short-term storage options for CECS are limited. Advantages of storage include: flexibility in scheduling surgery, reserve sheets for repeat operations, more opportunity for quality control, and improved transportation to allow wider distribution. Studies on storage of CECS have thus far focused on cryopreservation, whereas refrigeration is a convenient method commonly used for whole skin graft storage in burns clinics. It has been shown that preservation of viable cells using these methods is variable. This study evaluated the effect of different temperatures spanning 4°C to 37°C, on the cell viability, morphology, proliferation and metabolic status of CECS stored over a two week period in a xenobiotic–free system. Compared to non-stored control, best cell viability was obtained at 24°C (95.2±9.9%); reduced cell viability, at approximately 60%, was demonstrated at several of the temperatures (12°C, 28°C, 32°C and 37°C). Metabolic activity was significantly higher between 24°C and 37°C, where glucose, lactate, lactate/glucose ratios, and oxygen tension indicated increased activation of the glycolytic pathway under aerobic conditions. Preservation of morphology as shown by phase contrast and scanning electron micrographs was best at 12°C and 16°C. PCNA immunocytochemistry indicated that only 12°C and 20°C allowed maintenance of proliferative function at a similar level to non-stored control. In conclusion, results indicate that 12°C and 24°C merit further investigation as the prospective optimum temperature for short-term storage of cultured epidermal cell sheets.

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“…In the search for simpler and less expensive methods, some authors have described the cryopreservation of skin, but with the preservation period of 30 days 15,17 . In addition, other authors advocated the use of glycerol-preserved skin allografts.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, the cryopreservation of cultured epithelium has been a constant search topic in many centers worldwide 3,5,[13][14][15] .…”

Section: Introductionmentioning

confidence: 99%

An outcome analysis and long-term viability of cryopreserved cultured epidermal allografts: assessment of the conservation of transplantable human skin allografts

et al. 2013

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PURPOSE:To assess the viability of cultured epithelium and preserved by freezing for periods varying from one month to one year. METHODS:Samples of cultured epithelium were incubated in cryoprotectant medium (Group A), packed in aluminum envelopes and packed in polystyrene boxes. The boxes were subjected to a temperature of-70 o C. After freezing for a period of time ranging from one to 12 months, cultured epithelial samples were assessed for their viability by vital staining (Trypan blue) and metabolic analysis based on glucose consumption and lactate production. Samples of not frozen cultured epithelium (Group B) were also tested for viability and the results obtained were used as comparison parameter for the variation of viability. RESULTS:Statistical analysis between the group A and B indicate that the mean age of the donors (p=0.51) and the culture time (p=1.18) showed no statistical difference. In 30 days we obtained 37% of the original viability of cultured epithelium, 25% at six months and one year, less than 15%. This trend was confirmed statistically with a reduction of approximately 1.8% of the original viability epithelium cultured every 30 days of storage. In the analysis by lactate production, similar results were observed. In the analysis by the glucose consumption results were not significant. The viability indices show statistically significant difference between the group A and B (p<0.0001). CONCLUSIONS:Although cryopreserved cultured epithelium showed significant reduction of viability, all samples remained viable. It was also found that the viability of cryopreserved cultured epithelial decreased as a function of storage time.

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Skin Banking: A Simple Method for Cryopreservation of Split-Thickness Skin and Cultured Human Epidermal Keratinocytes

Cited by 16 publications

References 0 publications

Effectiveness of collagen/oxidised regenerated cellulose/silver‐containing composite wound dressing for the treatment of medium‐depth split‐thickness skin graft donor site wounds in multi‐morbid patients: a prospective, non‐comparative, single‐centre study

Effectiveness of collagen/oxidised regenerated cellulose/silver‐containing composite wound dressing for the treatment of medium‐depth split‐thickness skin graft donor site wounds in multi‐morbid patients: a prospective, non‐comparative, single‐centre study

Effect of Storage Temperature on Cultured Epidermal Cell Sheets Stored in Xenobiotic-Free Medium

An outcome analysis and long-term viability of cryopreserved cultured epidermal allografts: assessment of the conservation of transplantable human skin allografts

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