A proprietary GMP human platelet lysate for the expansion of dermal fibroblasts for clinical applications
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ABSTRACT:
PROVIDER: PRJNA613196 | ENA |
REPOSITORIES: ENA
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Project description: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.
2020-10-01 | GSE147146 | GEO
Project description:Mesenchymal stromal cells in platelet lysate versus fetal calf serum
| PRJNA347630 | ENA
Project description:Culture medium of mesenchymal stromal cells (MSCs) is usually supplemented with either human platelet lysate (HPL) or fetal calf serum (FCS). Many studies have demonstrated that proliferation and cellular morphology is influenced by these additives – hence they may favor outgrowth of specific subpopulations, thereby affecting the heterogeneous composition of MSCs. We have isolated and expanded human bone marrow derived MSCs in parallel with HPL or FCS for two passages. In HPL the proliferation was significantly higher and cells reflected more spindle-shaped morphology. Pairwise comparisons of gene expression profiles (Affymetrix HTA 2.0) revealed only moderate differences. When we apply a fold change >1.5 and limma-adjusted P-value of <0.05, only 69 transcripts were differentially expressed. These results indicate that there is no systematic bias for specific subpopulations of MSCs by using either HPL or FCS.
2017-08-11 | GSE87796 | GEO
Project description:Culture medium of mesenchymal stromal cells (MSCs) is usually supplemented with either human platelet lysate (HPL) or fetal calf serum (FCS). Many studies have demonstrated that proliferation and cellular morphology is influenced by these additives – hence they may favor outgrowth of specific subpopulations, thereby affecting the heterogeneous composition of MSCs. We have isolated and expanded human bone marrow derived MSCs in parallel with HPL or FCS for two passages. In HPL the proliferation was significantly higher and cells reflected more spindle-shaped morphology. In contrast, global DNA-methylation profiles did not reveal any significant differences. None of the CpGs revealed significant differences between MSCs that were cultured in HPL versus FCS if the analysis was corrected for multiple testing (limma-adjusted P-value of <0.05). These results indicate that there is no systematic bias for specific subpopulations of MSCs by using either HPL or FCS.
2017-08-11 | GSE87797 | GEO
Project description:The profiles of transcripts from the V. vulnificus grown under normal and high c-di-GMP conditions were compared by using a V. vulnificus whole-genome microarray Two-condition experiment, normal c-di-GMP condition vs. high c-di-GMP condition. Biological replicates: 3 control, 3 experimental, independently grown and harvested. One replicate per array. For transcriptome analysis, the V. vulnificus whole genome TwinChip, manufactured and kindly provided by the 21C Frontier Microbial Genomics and Applications Center (Daejeon, South Korea), was used.
2013-10-22 | E-GEOD-51459 | biostudies-arrayexpress
Project description:LC-MS/MS analysis of Control, PAR1-stimulated and PAR4-stimulated platelet lysate digests. Control lysate digest peptides were labeled with "light", PAR1 lysate digest peptides were labeled with "intermediate" and PAR4 lysate digest peptides were labeled with "high" dimethyl labeled. Labeled digests were mixed in a 1:1:1 ratio, separated by SCX and the resulting (phosphopeptide-enriched) peptide fractions were analyzed by LC-MS/MS.
2016-05-11 | PXD000228 | Pride
Project description:Stem cells from human exfoliated deciduous teeth (SHED) can be used as a potential clinical material. But the use of xenogeneic ingredients, especially fetal bovine serum, will increase the risk of zoonotic disease transmission during the use of animal-derived products. Human-platelet-lysate (hPL) was used in human cell expansion with reliability in clinical applications. This study aimed to explore the feasibility and effect of HPL in the cultivation of SHED. We use proteomics to interpret the impact of HPL on the protein profile of SHED. Propagation, differentiation, qPCR and immunofluorescence were used to detect the effect of HPL on SHED.
2022-05-19 | PXD026687 | Pride
Project description:Osteogenic transdifferentiation of primary human fibroblasts to osteoblast-like cells with human platelet lysate
| PRJNA851430 | ENA
Project description:Inherited bone disorders account for about 10% of documented Mendelian disorders and are associated with high financial burden. Their study requires osteoblasts which play a critical role in regulating the development and maintenance of bone tissue. However, bone tissue is not always available from patients. We developed a highly efficient platelet lysate-based approach to directly transdifferentiate skin-derived human fibroblasts to osteoblast-like cells. We extensively characterized our in vitro model by examining the expression of osteoblast-specific markers during the transdifferentiation process both at the mRNA and protein level. The transdifferentiated osteoblast-like cells showed significantly increased expression of a panel of osteogenic markers. Mineral deposition and ALP activity were also shown, confirming their osteogenic properties. RNA-seq analysis allowed the global study of changes in the transcriptome of the transdifferentiated cells. The transdifferentiated cells clustered separately from the fibroblasts with regard to the significantly upregulated genes indicating a distinct transcriptome profile; transdifferentiated osteoblasts also showed significant enrichment in gene expression related to skeletal development and bone mineralization. Our presented in vitro model may potentially contribute to the prospect of studying osteoblast-dependent disorders in patient-derived cells.
2022-06-24 | GSE206594 | GEO
Project description:Coccolithophore viral lysate Raw sequence reads
| PRJNA958194 | ENA