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In vitro dedifferentiation of melanocytes from adult epidermis.

Kormos B, Belso N, Bebes A, Szabad G, Bacsa S, Széll M, Kemény L, Bata-Csörgo Z - PLoS ONE (2011)

Bottom Line: In Mel-mix medium the cultured melanocytes are bipolar, unpigmented and highly proliferative.TRP-1 mRNA expression was induced in dedifferentiated melanocytes by UV-B irradiated keratinocyte supernatants, however direct UV-B irradiation of the cells resulted in further decrease of TRP-1 mRNA expression.These dedifferentiated, easily accessible cultured melanocytes provide a good model for studying melanocyte differentiation and possibly transdifferentiation.

View Article: PubMed Central - PubMed

Affiliation: Dermatological Research Group of the Hungarian Academy of Sciences, Szeged, Hungary. kormosbetti@mail.derma.szote.u-szeged.hu

ABSTRACT
In previous work we described a novel culture technique using a cholera toxin and PMA-free medium (Mel-mix) for obtaining pure melanocyte cultures from human adult epidermis. In Mel-mix medium the cultured melanocytes are bipolar, unpigmented and highly proliferative. Further characterization of the cultured melanocytes revealed the disappearance of c-Kit and TRP-1 and induction of nestin expression, indicating that melanocytes dedifferentiated in this in vitro culture. Cholera toxin and PMA were able to induce c-Kit and TRP-1 protein expressions in the cells, reversing dedifferentiation. TRP-1 mRNA expression was induced in dedifferentiated melanocytes by UV-B irradiated keratinocyte supernatants, however direct UV-B irradiation of the cells resulted in further decrease of TRP-1 mRNA expression. These dedifferentiated, easily accessible cultured melanocytes provide a good model for studying melanocyte differentiation and possibly transdifferentiation. Because melanocytes in Mel-mix medium can be cultured with human serum as the only supplement, this culture system is also suitable for autologous cell transplantation.

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Melanocyte cultures contain about equal number of senescent cells regardless of culture conditions.β-galactosidase assay was performed to detect senescent cells (blue cells) in the cultures. 3rd passage melanocytes cultured in Mel-mix medium (A), 3rd passage melanocytes cultured in M254 medium (B), 8th passage melanocytes cultured in Mel-mix medium (C), 8th passage melanocytes cultured in M254 medium (D). Bar: 100 µm.
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pone-0017197-g005: Melanocyte cultures contain about equal number of senescent cells regardless of culture conditions.β-galactosidase assay was performed to detect senescent cells (blue cells) in the cultures. 3rd passage melanocytes cultured in Mel-mix medium (A), 3rd passage melanocytes cultured in M254 medium (B), 8th passage melanocytes cultured in Mel-mix medium (C), 8th passage melanocytes cultured in M254 medium (D). Bar: 100 µm.

Mentions: Senescence-associated β-galactosidase assay was performed on both types of cultures when cells were in 3rd and 8th passages. Blue colored senescent cells were present in comparable numbers in both types of cultures regardless of the applied media. In 3rd passage cultures 50% of the cells were β-galactosidase+ in Mel-mix and 57% in M254, in 8th passage cultures 57% β-galactosidase+ cells were in Mel-mix and 48% in M254 (Figure 5). Blue cells appeared flat without dendrites in cultures, irrespective of the used media.


In vitro dedifferentiation of melanocytes from adult epidermis.

Kormos B, Belso N, Bebes A, Szabad G, Bacsa S, Széll M, Kemény L, Bata-Csörgo Z - PLoS ONE (2011)

Melanocyte cultures contain about equal number of senescent cells regardless of culture conditions.β-galactosidase assay was performed to detect senescent cells (blue cells) in the cultures. 3rd passage melanocytes cultured in Mel-mix medium (A), 3rd passage melanocytes cultured in M254 medium (B), 8th passage melanocytes cultured in Mel-mix medium (C), 8th passage melanocytes cultured in M254 medium (D). Bar: 100 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3044174&req=5

pone-0017197-g005: Melanocyte cultures contain about equal number of senescent cells regardless of culture conditions.β-galactosidase assay was performed to detect senescent cells (blue cells) in the cultures. 3rd passage melanocytes cultured in Mel-mix medium (A), 3rd passage melanocytes cultured in M254 medium (B), 8th passage melanocytes cultured in Mel-mix medium (C), 8th passage melanocytes cultured in M254 medium (D). Bar: 100 µm.
Mentions: Senescence-associated β-galactosidase assay was performed on both types of cultures when cells were in 3rd and 8th passages. Blue colored senescent cells were present in comparable numbers in both types of cultures regardless of the applied media. In 3rd passage cultures 50% of the cells were β-galactosidase+ in Mel-mix and 57% in M254, in 8th passage cultures 57% β-galactosidase+ cells were in Mel-mix and 48% in M254 (Figure 5). Blue cells appeared flat without dendrites in cultures, irrespective of the used media.

Bottom Line: In Mel-mix medium the cultured melanocytes are bipolar, unpigmented and highly proliferative.TRP-1 mRNA expression was induced in dedifferentiated melanocytes by UV-B irradiated keratinocyte supernatants, however direct UV-B irradiation of the cells resulted in further decrease of TRP-1 mRNA expression.These dedifferentiated, easily accessible cultured melanocytes provide a good model for studying melanocyte differentiation and possibly transdifferentiation.

View Article: PubMed Central - PubMed

Affiliation: Dermatological Research Group of the Hungarian Academy of Sciences, Szeged, Hungary. kormosbetti@mail.derma.szote.u-szeged.hu

ABSTRACT
In previous work we described a novel culture technique using a cholera toxin and PMA-free medium (Mel-mix) for obtaining pure melanocyte cultures from human adult epidermis. In Mel-mix medium the cultured melanocytes are bipolar, unpigmented and highly proliferative. Further characterization of the cultured melanocytes revealed the disappearance of c-Kit and TRP-1 and induction of nestin expression, indicating that melanocytes dedifferentiated in this in vitro culture. Cholera toxin and PMA were able to induce c-Kit and TRP-1 protein expressions in the cells, reversing dedifferentiation. TRP-1 mRNA expression was induced in dedifferentiated melanocytes by UV-B irradiated keratinocyte supernatants, however direct UV-B irradiation of the cells resulted in further decrease of TRP-1 mRNA expression. These dedifferentiated, easily accessible cultured melanocytes provide a good model for studying melanocyte differentiation and possibly transdifferentiation. Because melanocytes in Mel-mix medium can be cultured with human serum as the only supplement, this culture system is also suitable for autologous cell transplantation.

Show MeSH
Related in: MedlinePlus