The Use of Cell-penetrating Peptide for Delivery of Recombinant Transcription Factor DNA into Primary Human Fibroblast
Background: Reprogrammed cell therapy has not been applied for clinical purposes due to the malignancy issue. The aim of this study was to design the recombinant vector of the transcription factors and analyze the effectiveness of cell-penetrating peptide delivering system for human primary fibrobla...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Cell and BioPharmaceutical Institute
2023-03-01
|
| Series: | MCBS (Molecular and Cellular Biomedical Sciences) |
| Online Access: | https://cellbiopharm.com/ojs/index.php/MCBS/article/view/279 |
| _version_ | 1797858612858585088 |
|---|---|
| author | Melinda Remelia Budiman Bela Silvia Tri Widyaningtyas Radiana Dhewayani Antarianto Nuzli Fahdia Mazfufah Jeanne Adiwinata Pawitan |
| author_facet | Melinda Remelia Budiman Bela Silvia Tri Widyaningtyas Radiana Dhewayani Antarianto Nuzli Fahdia Mazfufah Jeanne Adiwinata Pawitan |
| author_sort | Melinda Remelia |
| collection | DOAJ |
| description | Background: Reprogrammed cell therapy has not been applied for clinical purposes due to the malignancy issue. The aim of this study was to design the recombinant vector of the transcription factors and analyze the effectiveness of cell-penetrating peptide delivering system for human primary fibroblast transfection to avoid the malignancy issue.
Materials and methods: The constructions of CCAT/enhancer binding protein alpha (CEBPA), hepatocyte nuclear factor 4 alpha (HNF4A), nuclear receptor subfamily 1 group I member 2 (NR1I2) were confirmed with DNA digestion and sequencing. Breast reduction (BRED) and palate (PAL) tissue were used as human primary fibroblast sources. The transcription factors were delivered into BRED and PAL with recombination of avian leukosis sarcoma virus (ALSV), human immunodeficiency virus (HIV) matrix, and regulator of expression of virion proteins (Rev) (ALMR), tagged with enhanced green fluorescence protein (eGFP). Post-transfection cells were then cultivated with optimized medium. Gene expression was measured with quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR).
Results: Gene expression levels of CEBPA, HNF4A, NR1I2, glutamate-ammonia ligase (GLUL), albumin (ALB), and cytochrome P450 (CYP) were increased. Transfection with ALMR, which were more efficient in BRED than PAL fibroblasts may have the advantage in autologous cell therapy for elderly patients.
Conclusion: Transfection of transcription factors to human primary fibroblast may be performed by using constructions of plasmid as designed in this study.
Keywords: recombinant plasmid, hepatocyte-like cells, primary fibroblasts, recombinant peptide, cell reprogramming, autologous cells therapy |
| first_indexed | 2024-04-09T21:16:09Z |
| format | Article |
| id | doaj.art-465fd28537a14e35ac6281b30a1a0b0b |
| institution | Directory Open Access Journal |
| issn | 2527-4384 2527-3442 |
| language | English |
| last_indexed | 2024-04-09T21:16:09Z |
| publishDate | 2023-03-01 |
| publisher | Cell and BioPharmaceutical Institute |
| record_format | Article |
| series | MCBS (Molecular and Cellular Biomedical Sciences) |
| spelling | doaj.art-465fd28537a14e35ac6281b30a1a0b0b2023-03-28T07:46:24ZengCell and BioPharmaceutical InstituteMCBS (Molecular and Cellular Biomedical Sciences)2527-43842527-34422023-03-0171283710.21705/mcbs.v7i1.279105The Use of Cell-penetrating Peptide for Delivery of Recombinant Transcription Factor DNA into Primary Human FibroblastMelinda Remelia0Budiman Bela1Silvia Tri Widyaningtyas2Radiana Dhewayani Antarianto3Nuzli Fahdia Mazfufah4Jeanne Adiwinata Pawitan5Department of Histology, Faculty of Medicine, Universitas Kristen Indonesia, JakartaDoctoral Program, Biomedical Science, Faculty of Medicine, Universitas Indonesia, JakartaVirology and Cancer Pathobiology Research Center, Faculty of Medicine, Universitas Indonesia/Cipto Mangunkusumo National Central General Hospital, JakartaDoctoral Program, Biomedical Science, Faculty of Medicine, Universitas Indonesia, JakartaStem Cell Research and Tissue Engineering - Indonesia Medical Education and Research Institute, Faculty of Medicine, Universitas Indonesia, JakartaDoctoral Program, Biomedical Science, Faculty of Medicine, Universitas Indonesia, JakartaBackground: Reprogrammed cell therapy has not been applied for clinical purposes due to the malignancy issue. The aim of this study was to design the recombinant vector of the transcription factors and analyze the effectiveness of cell-penetrating peptide delivering system for human primary fibroblast transfection to avoid the malignancy issue. Materials and methods: The constructions of CCAT/enhancer binding protein alpha (CEBPA), hepatocyte nuclear factor 4 alpha (HNF4A), nuclear receptor subfamily 1 group I member 2 (NR1I2) were confirmed with DNA digestion and sequencing. Breast reduction (BRED) and palate (PAL) tissue were used as human primary fibroblast sources. The transcription factors were delivered into BRED and PAL with recombination of avian leukosis sarcoma virus (ALSV), human immunodeficiency virus (HIV) matrix, and regulator of expression of virion proteins (Rev) (ALMR), tagged with enhanced green fluorescence protein (eGFP). Post-transfection cells were then cultivated with optimized medium. Gene expression was measured with quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Results: Gene expression levels of CEBPA, HNF4A, NR1I2, glutamate-ammonia ligase (GLUL), albumin (ALB), and cytochrome P450 (CYP) were increased. Transfection with ALMR, which were more efficient in BRED than PAL fibroblasts may have the advantage in autologous cell therapy for elderly patients. Conclusion: Transfection of transcription factors to human primary fibroblast may be performed by using constructions of plasmid as designed in this study. Keywords: recombinant plasmid, hepatocyte-like cells, primary fibroblasts, recombinant peptide, cell reprogramming, autologous cells therapyhttps://cellbiopharm.com/ojs/index.php/MCBS/article/view/279 |
| spellingShingle | Melinda Remelia Budiman Bela Silvia Tri Widyaningtyas Radiana Dhewayani Antarianto Nuzli Fahdia Mazfufah Jeanne Adiwinata Pawitan The Use of Cell-penetrating Peptide for Delivery of Recombinant Transcription Factor DNA into Primary Human Fibroblast MCBS (Molecular and Cellular Biomedical Sciences) |
| title | The Use of Cell-penetrating Peptide for Delivery of Recombinant Transcription Factor DNA into Primary Human Fibroblast |
| title_full | The Use of Cell-penetrating Peptide for Delivery of Recombinant Transcription Factor DNA into Primary Human Fibroblast |
| title_fullStr | The Use of Cell-penetrating Peptide for Delivery of Recombinant Transcription Factor DNA into Primary Human Fibroblast |
| title_full_unstemmed | The Use of Cell-penetrating Peptide for Delivery of Recombinant Transcription Factor DNA into Primary Human Fibroblast |
| title_short | The Use of Cell-penetrating Peptide for Delivery of Recombinant Transcription Factor DNA into Primary Human Fibroblast |
| title_sort | use of cell penetrating peptide for delivery of recombinant transcription factor dna into primary human fibroblast |
| url | https://cellbiopharm.com/ojs/index.php/MCBS/article/view/279 |
| work_keys_str_mv | AT melindaremelia theuseofcellpenetratingpeptidefordeliveryofrecombinanttranscriptionfactordnaintoprimaryhumanfibroblast AT budimanbela theuseofcellpenetratingpeptidefordeliveryofrecombinanttranscriptionfactordnaintoprimaryhumanfibroblast AT silviatriwidyaningtyas theuseofcellpenetratingpeptidefordeliveryofrecombinanttranscriptionfactordnaintoprimaryhumanfibroblast AT radianadhewayaniantarianto theuseofcellpenetratingpeptidefordeliveryofrecombinanttranscriptionfactordnaintoprimaryhumanfibroblast AT nuzlifahdiamazfufah theuseofcellpenetratingpeptidefordeliveryofrecombinanttranscriptionfactordnaintoprimaryhumanfibroblast AT jeanneadiwinatapawitan theuseofcellpenetratingpeptidefordeliveryofrecombinanttranscriptionfactordnaintoprimaryhumanfibroblast AT melindaremelia useofcellpenetratingpeptidefordeliveryofrecombinanttranscriptionfactordnaintoprimaryhumanfibroblast AT budimanbela useofcellpenetratingpeptidefordeliveryofrecombinanttranscriptionfactordnaintoprimaryhumanfibroblast AT silviatriwidyaningtyas useofcellpenetratingpeptidefordeliveryofrecombinanttranscriptionfactordnaintoprimaryhumanfibroblast AT radianadhewayaniantarianto useofcellpenetratingpeptidefordeliveryofrecombinanttranscriptionfactordnaintoprimaryhumanfibroblast AT nuzlifahdiamazfufah useofcellpenetratingpeptidefordeliveryofrecombinanttranscriptionfactordnaintoprimaryhumanfibroblast AT jeanneadiwinatapawitan useofcellpenetratingpeptidefordeliveryofrecombinanttranscriptionfactordnaintoprimaryhumanfibroblast |