Keloid explant culture: a model for keloid fibroblasts isolation and cultivation based on the biological differences of its specific regions.
In vitro studies with keloid fibroblasts frequently present contradictory results. This may occur because keloids present distinct genotypic and phenotypic characteristics in its different regions, such as the peripheral region in relation to the central region. We suggest an explant model for keloid fibroblasts harvesting, standardising the initial processing of keloid samples to obtain fragments from different regions, considering its biological differences, for primary cell culture. The different keloid regions were delimited and fragments were obtained using a 3-mm diameter punch. To remove fragments from the periphery, the punch was placed in one longitudinal line extremity, respecting the lesion borders. For the central region, it was placed in the intersection of lines at the level of the largest longitudinal and transversal axes, the other fragments being removed centrifugally in relation to the first one. Primary fibroblast culture was carried out by explant. Flow cytometry analysis showed cell cycle differences between the groups, confirming its different origins and biological characteristics. In conclusion, our proposed model proved itself efficient for keloid fibroblast isolation from specific regions and cultivation. Its simplicity and ease of execution may turn it into an important tool for studying the characteristics of the different keloid-derived fibroblasts in culture.
Tucci-Viegas
Vanina Monique
VM
Laboratory of Cell Culture, Plastic Surgery Division, Universidade Federal de São Paulo, São Paulo, Brazil. vanina monique@terra.com.br
Hochman
Bernardo
B
França
Jerônimo P
JP
Ferreira
Lydia M
LM
D000293:Adolescent; D000328:Adult; D005260:Female; D005347:Fibroblasts; D006801:Humans; D007627:Keloid; D013048:Specimen Handling / Q000379:methods*; D046509:Tissue Culture Techniques; D055815:Young Adult