Long term effect of cryopreservation on primary human skin cells

M. F. Ishak, M. Manira, Min Hwei Ng, B. Khairul, L. Gargy, B. S. Aminuddin, Ruszymah Idrus

Research output: Contribution to journalArticle

Abstract

Cryopreservation is essential for tissue engineering and regenerative medicine. This study was carried out to assess the effect of cryopreservation on skin cells and evaluate the performance of cells after 12 months of cryopreservation. Redundant skin tissue samples were obtained from surgery with consent from patients. The tissue was cleaned, processed and cultured until passage 3. Upon confluency, cells were trypsinised and total cell yield and viability were determined before and after being cryopreserved. Sterility and immunocytochemistry analysis for collagen type I (Col-1) and cytokeratin 14 (CK14) antibodies were also performed on cells cryopreserved for one, three, six and twelve months. There is no significant difference in growth rates for cryopreserved cells for 1 to 12 months, except for fibroblasts at 6 months. Cell viability for both keratinocytes and fibroblasts decreased with time (65%± 3.5% - 89%± 4.5%). Sterility testing showed no contamination after 12 months of cryopreservation. Immunocytochemistry analysis showed positive expression for CK14 (keratinocytes) and Col -1 (fibroblasts) after 12 months of cryopreservation. Morphologically, keratinocytes and fibroblasts were able to retain its phenotype. The loss in viability is consistent in all samples and possibly due to thermal-cycling effect. Immunocytochemistry and consistent cell growth analysis showed that keratinocytes and fibroblasts were able to retain their characteristics in cryopreservation condition. These preliminary findings show that primary skin cells can be stored via cryopreservation and still retain their characteristics. However, further investigations using longer periods of cryopreservation (24 months, 48 months) should be conducted.

Translated title of the contributionLong term effect of cryopreservation on primary human skin cells
Original languageMalay
Pages (from-to)137-144
Number of pages8
JournalSains Malaysiana
Volume48
Issue number1
DOIs
Publication statusPublished - 1 Jan 2019

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Cryopreservation
Skin
Keratinocytes
Fibroblasts
Keratin-14
Immunohistochemistry
Infertility
Cell Survival
Regenerative Medicine
Tissue Engineering
Growth
Collagen Type I
Hot Temperature
Phenotype
Antibodies

Keywords

    ASJC Scopus subject areas

    • General

    Cite this

    Kesan jangka panjang pengawetan krio pada sel kulit primer manusia. / Ishak, M. F.; Manira, M.; Ng, Min Hwei; Khairul, B.; Gargy, L.; Aminuddin, B. S.; Idrus, Ruszymah.

    In: Sains Malaysiana, Vol. 48, No. 1, 01.01.2019, p. 137-144.

    Research output: Contribution to journalArticle

    Ishak, MF, Manira, M, Ng, MH, Khairul, B, Gargy, L, Aminuddin, BS & Idrus, R 2019, 'Kesan jangka panjang pengawetan krio pada sel kulit primer manusia' Sains Malaysiana, vol. 48, no. 1, pp. 137-144. https://doi.org/10.17576/jsm-2019-4801-16
    Ishak MF, Manira M, Ng MH, Khairul B, Gargy L, Aminuddin BS et al. Kesan jangka panjang pengawetan krio pada sel kulit primer manusia. Sains Malaysiana. 2019 Jan 1;48(1):137-144. https://doi.org/10.17576/jsm-2019-4801-16
    Ishak, M. F. ; Manira, M. ; Ng, Min Hwei ; Khairul, B. ; Gargy, L. ; Aminuddin, B. S. ; Idrus, Ruszymah. / Kesan jangka panjang pengawetan krio pada sel kulit primer manusia. In: Sains Malaysiana. 2019 ; Vol. 48, No. 1. pp. 137-144.
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    abstract = "Cryopreservation is essential for tissue engineering and regenerative medicine. This study was carried out to assess the effect of cryopreservation on skin cells and evaluate the performance of cells after 12 months of cryopreservation. Redundant skin tissue samples were obtained from surgery with consent from patients. The tissue was cleaned, processed and cultured until passage 3. Upon confluency, cells were trypsinised and total cell yield and viability were determined before and after being cryopreserved. Sterility and immunocytochemistry analysis for collagen type I (Col-1) and cytokeratin 14 (CK14) antibodies were also performed on cells cryopreserved for one, three, six and twelve months. There is no significant difference in growth rates for cryopreserved cells for 1 to 12 months, except for fibroblasts at 6 months. Cell viability for both keratinocytes and fibroblasts decreased with time (65{\%}± 3.5{\%} - 89{\%}± 4.5{\%}). Sterility testing showed no contamination after 12 months of cryopreservation. Immunocytochemistry analysis showed positive expression for CK14 (keratinocytes) and Col -1 (fibroblasts) after 12 months of cryopreservation. Morphologically, keratinocytes and fibroblasts were able to retain its phenotype. The loss in viability is consistent in all samples and possibly due to thermal-cycling effect. Immunocytochemistry and consistent cell growth analysis showed that keratinocytes and fibroblasts were able to retain their characteristics in cryopreservation condition. These preliminary findings show that primary skin cells can be stored via cryopreservation and still retain their characteristics. However, further investigations using longer periods of cryopreservation (24 months, 48 months) should be conducted.",
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