Long Term Effect of Cryopreservation on Primary Human Skin Cells

Ishak, Mohamad Fikeri; Maarof, Manira; Ng, Min Hwei; Khairul, B; Gargy, L; Aminuddin, B S; Idrus, Ruszymah Haji

doi:10.17576/jsm-2019-4801-16

Cited by 3 publications

(1 citation statement)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cryogenic preservation can maintain fibroblast cells for very long periods [9]. Subcultures with 3 or 4 passages should be used for cryogenic preservation.…”

Section: Freezing and Thawing Fibroblast Cellsmentioning

confidence: 99%

Simple Method for Establishing Primary Leporidae Skin Fibroblast Cultures

Santos

Carvalho

Almeida

et al. 2021

Cells

View full text Add to dashboard Cite

Commercial hare and rabbit immortalized cell lines are extremely limited regarding the many species within the lagomorpha order. To overcome this limitation, researchers and technicians must establish primary cell cultures derived from biopsies or embryos. Among all cell types, fibroblasts are plastic and resilient cells, highly convenient for clinical and fundamental research but also for diagnosis, particularly for viral isolation. Here, we describe a fast and cheap method to produce primary fibroblast cell cultures from leporid species, using dispase II, a protease that allows dermal–epidermal separation, followed by a simple enzymatic digestion with trypsin. This method allows for the establishment of an in vitro cell culture system with an excellent viability yield and purity level higher than 85% and enables the maintenance and even immortalization of leporid fibroblastic cells derived from tissues already differentiated.

show abstract

“…Cryogenic preservation can maintain fibroblast cells for very long periods [9]. Subcultures with 3 or 4 passages should be used for cryogenic preservation.…”

Section: Freezing and Thawing Fibroblast Cellsmentioning

confidence: 99%

Simple Method for Establishing Primary Leporidae Skin Fibroblast Cultures

Santos

Carvalho

Almeida

et al. 2021

Cells

View full text Add to dashboard Cite

show abstract

Optimisation of cryopreservation conditions, including storage duration and revival methods, for the viability of human primary cells

Mohamed,

Sundar,

Ridwan

et al. 2024

BMC Mol and Cell Biol

View full text Add to dashboard Cite

Background Cryopreservation is a crucial procedure for safeguarding cells or other biological constructs, showcasing considerable potential for applications in tissue engineering and regenerative medicine. Aims This study aimed to evaluate the effectiveness of different cryopreservation conditions on human cells viability. Methods A set of cryopreserved data from Department of Tissue Engineering and Regenerative Medicine (DTERM) cell bank were analyse for cells attachment after 24 h being revived. The revived cells were analysed based on different cryopreservation conditions which includes cell types (skin keratinocytes and fibroblasts, respiratory epithelial, bone marrow mesenchymal stem cell (MSC); cryo mediums (FBS + 10% DMSO; commercial medium); storage durations (0 to > 24 months) and locations (tank 1–2; box 1–5), and revival methods (direct; indirect methods). Human dermal fibroblasts (HDF) were then cultured, cryopreserved in different cryo mediums (HPL + 10% DMSO; FBS + 10% DMSO; Cryostor) and stored for 1 and 3 months. The HDFs were revived using either direct or indirect method and cell number, viability and protein expression analysis were compared. Results In the analysis cell cryopreserved data; fibroblast cells; FBS + 10% DMSO cryo medium; storage duration of 0–6 months; direct cell revival; storage in vapor phase of cryo tank; had the highest number of vials with optimal cell attachment after 24 h revived. HDFs cryopreserved in FBS + 10% DMSO for 1 and 3 months with both revival methods, showed optimal live cell numbers and viability above 80%, higher than other cryo medium groups. Morphologically, the fibroblasts were able to retain their phenotype with positive expression of Ki67 and Col-1. HDFs cryopreserved in FBS + 10% DMSO at 3 months showed significantly higher expression of Ki67 (97.3% ± 4.62) with the indirect revival method, while Col-1 expression (100%) was significantly higher at both 1 and 3 months compared to other groups. Conclusion In conclusion, fibroblasts were able to retain their characteristics after various cryopreservation conditions with a slight decrease in viability that may be due to the thermal-cycling effect. However, further investigation on the longer cryopreservation periods should be conducted for other types of cells and cryo mediums to achieve optimal cryopreservation outcomes.

show abstract

An Anhydrous Sodium Chloride Skin Preservation Model for Studies on Keratinocytes Grafting into the Wounds

et al. 2021

View full text Add to dashboard Cite

Background. Human skin is needed for covering large body areas lost by trauma. The shortcomings of contemporary methods of skin storage are limited preservation time and high immunogenicity if allogeneic. Methods. We investigated whether long-lasting skin preservation in anhydrous sodium chloride (NaCl) may be the source of keratinocytes (KCs) for transplantation. Dehydrated skin fragments were preserved for a time frame from 1 week to 12 months. Then, skin fragments were rehydrated, and KCs were isolated. The viability of KCs was assessed in viability/cytotoxicity test. NaCl-preserved KCs were cultured for 7 days and transplanted to the dorsum of SCID mice. Results. The morphology of NaCl-preserved KCs was unaltered. KCs from all epidermal layers could be identified. All grafts were accepted by the recipients. Transplanted KCs: synthesized keratins 10 and 16 expressed antigens specific for stem cells and transient-amplifying cells, and remained HLA-I-positive. Moreover, they expressed the proliferative marker PCNA. Cells isolated from transplants remained viable and produced enzymes. Conclusions. Transplantation of KCs obtained from human skin and stored in anhydrous NaCl may be considered for the closure of extensive skin wounds. The originality of this method consists of an effective storage procedure and easy preparation of keratinocytes for transplantation.

show abstract

Long Term Effect of Cryopreservation on Primary Human Skin Cells

Cited by 3 publications

References 31 publications

Simple Method for Establishing Primary Leporidae Skin Fibroblast Cultures

Simple Method for Establishing Primary Leporidae Skin Fibroblast Cultures

Optimisation of cryopreservation conditions, including storage duration and revival methods, for the viability of human primary cells

An Anhydrous Sodium Chloride Skin Preservation Model for Studies on Keratinocytes Grafting into the Wounds

Contact Info

Product

Resources

About