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Immortalization of Primary Keratinocytes and Its Application to Skin Research.

Choi M, Lee C - Biomol Ther (Seoul) (2015)

Bottom Line: Therefore, a number of immortalization attempts have been made with success to overcome this limitation.Then, various immortalization methods for the establishment of stable keratinocyte cell lines will be explained.Finally, its application to a three-dimensional skin culture system will be described.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Dongguk University-Seoul, Goyang 410-820, Republic of Korea.

ABSTRACT
As a major component of the epidermal tissue, a primary keratinocyte has served as an essential tool not only for the study of pathogenesis of skin-related diseases but also for the assessment of potential toxicities of various chemicals used in cosmetics. However, its short lifespan in ex vivo setting has been a great hurdle for many practical applications. Therefore, a number of immortalization attempts have been made with success to overcome this limitation. In order to understand the immortalization process of a primary keratinocyte, several key biological phenomena governing its lifespan will be reviewed first. Then, various immortalization methods for the establishment of stable keratinocyte cell lines will be explained. Finally, its application to a three-dimensional skin culture system will be described.

No MeSH data available.


Related in: MedlinePlus

Schematic diagram on how to immortalize primary keratinocytes by a retroviral transduction of HPV E6 and E7 oncogenes.
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f4-bt-23-391: Schematic diagram on how to immortalize primary keratinocytes by a retroviral transduction of HPV E6 and E7 oncogenes.

Mentions: For an efficient immortalization of primary keratinocytes, overexpression of HPV E6/E7 by retroviral transduction has been a method of choice so far thanks to its capacity to preserve normal cell-like properties in immortalized cells. A schematic diagram on how to use a retroviral transduction to induce immortalization of target cells was shown in Fig. 4. First, cDNA encoding HPV E6/E7 is cloned into a retroviral vector. Then, this cloned retroviral vector is transfected into 293T packaging cells together with other retroviral envelope or packaging vectors, followed by harvesting of retroviruses, which are encoding HPV E6/E7, in the supernatant. Then, primary keratinocytes, which are typically isolated from human foreskin tissues, are infected with these harvested retroviruses. Through a drug selection, immortalized keratinocytes, which are stably expressing HPV E6/E7, are isolated and propagated.


Immortalization of Primary Keratinocytes and Its Application to Skin Research.

Choi M, Lee C - Biomol Ther (Seoul) (2015)

Schematic diagram on how to immortalize primary keratinocytes by a retroviral transduction of HPV E6 and E7 oncogenes.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556197&req=5

f4-bt-23-391: Schematic diagram on how to immortalize primary keratinocytes by a retroviral transduction of HPV E6 and E7 oncogenes.
Mentions: For an efficient immortalization of primary keratinocytes, overexpression of HPV E6/E7 by retroviral transduction has been a method of choice so far thanks to its capacity to preserve normal cell-like properties in immortalized cells. A schematic diagram on how to use a retroviral transduction to induce immortalization of target cells was shown in Fig. 4. First, cDNA encoding HPV E6/E7 is cloned into a retroviral vector. Then, this cloned retroviral vector is transfected into 293T packaging cells together with other retroviral envelope or packaging vectors, followed by harvesting of retroviruses, which are encoding HPV E6/E7, in the supernatant. Then, primary keratinocytes, which are typically isolated from human foreskin tissues, are infected with these harvested retroviruses. Through a drug selection, immortalized keratinocytes, which are stably expressing HPV E6/E7, are isolated and propagated.

Bottom Line: Therefore, a number of immortalization attempts have been made with success to overcome this limitation.Then, various immortalization methods for the establishment of stable keratinocyte cell lines will be explained.Finally, its application to a three-dimensional skin culture system will be described.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmacy, Dongguk University-Seoul, Goyang 410-820, Republic of Korea.

ABSTRACT
As a major component of the epidermal tissue, a primary keratinocyte has served as an essential tool not only for the study of pathogenesis of skin-related diseases but also for the assessment of potential toxicities of various chemicals used in cosmetics. However, its short lifespan in ex vivo setting has been a great hurdle for many practical applications. Therefore, a number of immortalization attempts have been made with success to overcome this limitation. In order to understand the immortalization process of a primary keratinocyte, several key biological phenomena governing its lifespan will be reviewed first. Then, various immortalization methods for the establishment of stable keratinocyte cell lines will be explained. Finally, its application to a three-dimensional skin culture system will be described.

No MeSH data available.


Related in: MedlinePlus