|
| Status |
Public on Oct 01, 2020 |
| Title |
A proprietary GMP human platelet lysate for the expansion of dermal fibroblasts for clinical applications |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
|
| Summary |
Recent years have witnessed the introduction of ex vivo expanded dermal fibroblasts for several cell therapy and tissue-engineering applications, including the treatment of facial scars and burns, representing a promising cell type for regenerative medicine. We tested different in-house produced human platelet lysate (HPL) solutions against fetal bovine serum as supplements for in vitro fibroblast expansion by comparing cell yield, molecular marker expression, extracellular matrix (ECM) generation, genomic stability and global gene expression. Our in-house produced HPL supported fibroblast growth at levels similar to those for FBS and commercial HPL products and was superior to AB human serum. Cells grown in HPL maintained a fibroblast phenotype (VIM+, CD44+, CD13+, CD90+), ECM generation capacity (FN+, COL1+) and a normal karyotype, although gene expression profiling revealed changes related to cell metabolism, adhesion and cellular senescence. The HPL manufacturing process was validated within a GMP compliant system and the solution was stable at -80ºC and -20ºC for 2 years. Dermal fibroblasts expanded in vitro with HPL maintain a normal karyotype and expression of fibroblast markers, with only minor changes in their global gene expression profile. Our in-house produced GMP-HPL is an efficient, safe and economical cell culture supplement that can help increase the healthcare activity of blood transfusion centers through the re-use of transfusional plasma and platelets approaching their expiration date. Currently, our HPL solution is approved by the Spanish Agency of Medicines and Medical Devices and is being used in the manufacture of cell therapy products.
|
| |
|
| Overall design |
9 total samples were analyzed. Fibroblasts were cultured for 8 passages either with 10% Fetal Bovine Serum (FBS), 5% or 15% in-house HPL2 solution (n=3, respectively) as culture supplement. Biological replicates are included. Differentially expressed genes (DEGs) were obtained applying a threshold of ± 2 fold change as well as a false discovery rate p-value less than or equal to 0.05 (FDR-P val ≤ 0.05).
|
| Web link |
https://0-www-tandfonline-com.brum.beds.ac.uk/doi/full/10.1080/09537104.2020.1856356
|
| |
|
| Contributor(s) |
Santos-González M, Lopez-Navas L |
| Citation(s) |
33393414 |
| Submission date |
Mar 18, 2020 |
| Last update date |
Jan 07, 2021 |
| Contact name |
Luis Lopez Navas |
| Phone |
+34656523579
|
| Organization name |
Red Andaluza de diseño y traslación de Terapias Avanzadas
|
| Department |
Preclinical Research
|
| Street address |
c/ Americo Vespucio, 15, Ed. S-2
|
| City |
Sevilla |
| State/province |
Sevilla |
| ZIP/Postal code |
41092 |
| Country |
Spain |
| |
|
| Platforms (1) |
| GPL23159 |
[Clariom_S_Human] Affymetrix Clariom S Assay, Human (Includes Pico Assay) |
|
| Samples (9)
|
|
| Relations |
| BioProject |
PRJNA613196 |
Less...
More...