RT Journal Article
T1 Improvement of Cell Culture Methods for the Successful Generation of Human Keratinocyte Primary Cell Cultures Using EGF-Loaded Nanostructured Lipid Carriers
A1 Chato-Astrain, Jesús
A1 Sánchez-Porras, David
A1 García-García, Óscar Darío
A1 Vairo, Claudia
A1 Villar-Vidal, María
A1 Villullas, Silvia
A1 Sánchez-Montesinos, Indalecio
A1 Campos, Fernando
A1 Garzón, Ingrid
A1 Alaminos, Miguel
K1 EGF
K1 Nanoparticles
K1 NLC
K1 Keratinocytes
K1 Tissue engineering
K1 Cell culture
K1 Epidermal growth factor
K1 Factor de crecimiento epidérmico
K1 Nanopartículas
K1 Queratinocitos
K1 Ingeniería de tejidos
K1 Técnicas de cultivo de célula
AB Human skin keratinocyte primary cultures can be established from skin biopsies with culture media containing epithelial growth factor (EGF). Although current methods are efficient, optimization is required to accelerate the procedure and obtain these cultures in less time. In the present study, we evaluated the effect of novel formulations based on EGF-loaded nanostructured lipid carriers (NLC). First, biosafety of NLC containing recombinant human EGF (NLC-rhEGF) was verified in immortalized skin keratinocytes and cornea epithelial cells, and in two epithelial cancer cell lines, by quantifying free DNA released to the culture medium. Then we established primary cell cultures of human skin keratinocytes with basal culture media (BM) and BM supplemented with NLC-rhEGF, liquid EGF (L-rhEGF), or NLC alone (NLC-blank). The results showed that cells isolated by enzymatic digestion and cultured with or without a feeder layer had a similar growth rate regardless of the medium used. However, the explant technique showed higher efficiency when NLC-rhEGF culture medium was used, compared to BM, L-rhEGF, or NLC-blank. Gene expression analysis showed that NLC-rhEGF was able to increase EGFR gene expression, along with that of other genes related to cytokeratins, cell-cell junctions, and keratinocyte maturation and differentiation. In summary, these results support the use of NLC-rhEGF to improve the efficiency of explant-based methods in the efficient generation of human keratinocyte primary cell cultures for tissue engineering use.
PB MDPI
YR 2021
FD 2021-11-06
LK http://hdl.handle.net/10668/3494
UL http://hdl.handle.net/10668/3494
LA en
NO Chato-Astrain J, Sánchez-Porras D, García-García Ó.D, Vairo C, Villar-Vidal M, Villullas S, et al. Improvement of Cell Culture Methods for the Successful Generation of Human Keratinocyte Primary Cell Cultures Using EGF-Loaded Nanostructured Lipid Carriers. Biomedicines 2021, 9 (11), 1634.
DS RISalud
RD Apr 7, 2025