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Life
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Cell Culture and Genome Modification Technologies: from Biology to Biomedical...
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Cell Culture and Genome Modification Technologies: from Biology to Biomedical Applications

A special issue of Life (ISSN 2075-1729). This special issue belongs to the section "Cell Biology and Tissue Engineering".

Deadline for manuscript submissions: closed (31 August 2022) | Viewed by 9014

Special Issue Editors

Dr. Fabiana Fernandes Bressan

E-Mail Website
Guest Editor
Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, Pirassununga 13635-000, Brazil
Interests: developmental biology cellular; epigenetic reprogramming; stem cells; cell therapy and gene therapy; induced reprogramming; in vitro gametogenesis; cultivated meat
Special Issues, Collections and Topics in MDPI journals
Dr. Kelly Roballo

E-Mail Website
Guest Editor
1.Department of Cell Biology and Physiology, Edward via College of Osteopathic Medicine, Blacksburg, VA, USA
2.Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Tech, Blacksburg, VA, USA
Interests: stem cell; immunology; peripheral nerve injury
Dr. Naira Caroline Godoy Pieri

E-Mail Website
Assistant Guest Editor
Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, Pirassununga 05508-060, Brazil
Interests: pluripotent stem cells; germ cells; pigs and dogs
Dr. Lais Vicari de Figueiredo Pessôa

E-Mail Website
Assistant Guest Editor
Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, Pirassununga 05508-060, Brazil
Interests: cell line culture; plasmid cloning; cellular differentiation

Special Issue Information

Dear Colleagues,

Regenerative medicine is a rapidly ascending field supported mainly by the development of new biotechnologies, particularly the possibility of genetically modifying the inherited genome and epigenetically modifying the cell fates. Significant advances using cell culture and genome modification technologies have been reported to establish viable conditions and models that will enable the modulation of cell genetic information and phenotypes for practical use in medicine (for example, modeling genetic diseases, cancer, or fertility purposes). They may also provide beneficial technologies for agriculture (optimization of herds, climate issues, etc.). However, essential barriers related to the translation of protocols and results are still common, mainly concerning the establishment of cell lines, robust editing and differentiation protocols, the use of large-scale culture systems, and pre-clinical or clinical assessments. Therefore, the present Special Issue aims to present consolidated results and perspectives on the culture and manipulation of cells in vitro, aiming for basic and applied purposes.

Dr. Fabiana Fernandes Bressan
Dr. Kelly Roballo
Dr. Naira Caroline Godoy Pieri
Dr. Lais Vicari de Figueiredo Pessôa
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Life is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • regenerative medicine
  • translational medicine
  • cellular modeling
  • gene editing
  • genetic engineering

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Published Papers (3 papers)

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Research

10 pages, 5886 KiB  
Article
Influence of Cell Type in In Vitro Induced Reprogramming in Cattle
by Kaiana Recchia, Laís Vicari de Figueiredo Pessôa, Naira Caroline Godoy Pieri, Pedro Ratto Lisboa Pires and Fabiana Fernandes Bressan
Life 2022, 12(8), 1139; https://doi.org/10.3390/life12081139 - 28 Jul 2022
Cited by 2 | Viewed by 2051
Abstract
Induced pluripotent stem cells (iPSCs) have been considered an essential tool in stem cell research due to their potential to develop new therapies and technologies and answer essential questions about mammalian early development. An important step in generating iPSCs is selecting their precursor [...] Read more.
Induced pluripotent stem cells (iPSCs) have been considered an essential tool in stem cell research due to their potential to develop new therapies and technologies and answer essential questions about mammalian early development. An important step in generating iPSCs is selecting their precursor cell type, influencing the reprogramming efficiency and maintenance in culture. In this study, we aim to characterize bovine mesenchymal cells from adipose tissue (bAdMSCs) and fetal fibroblasts (bFFs) and to compare the reprogramming efficiency of these cells when induced to pluripotency. The cells were characterized by immunostaining (CD90, SSEA1, SSEA3, and SSEA4), induced differentiation in vitro, proliferation rates, and were subjected to cell reprogramming using the murine OSKM transcription factors. The bFFs presented morphological changes resembling pluripotent cells after reprogramming and culture with different supplementation, and putative iPSCs were characterized by immunostaining (OCT4, SOX2, NANOG, and AP). In the present study, we demonstrated that cell line origin and cellular proliferation rate are determining factors for reprogramming cells into pluripotency. The generation of biPSCs is a valuable tool to improve both translational medicine and animal production and to study the different supplements required to maintain the pluripotency of bovine cells in vitro. Full article
(This article belongs to the Special Issue Cell Culture and Genome Modification Technologies: from Biology to Biomedical Applications)
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13 pages, 2056 KiB  
Article
Cell Cycle Stage and DNA Repair Pathway Influence CRISPR/Cas9 Gene Editing Efficiency in Porcine Embryos
by Karina Gutierrez, Werner G. Glanzner, Mariana P. de Macedo, Vitor B. Rissi, Naomi Dicks, Rodrigo C. Bohrer, Hernan Baldassarre, Luis B. Agellon and Vilceu Bordignon
Life 2022, 12(2), 171; https://doi.org/10.3390/life12020171 - 25 Jan 2022
Cited by 3 | Viewed by 3369
Abstract
CRISPR/Cas9 technology is a powerful tool used for genome manipulation in different cell types and species. However, as with all new technologies, it still requires improvements. Different factors can affect CRISPR/Cas efficiency in zygotes, which influence the total cost and complexity for creating [...] Read more.
CRISPR/Cas9 technology is a powerful tool used for genome manipulation in different cell types and species. However, as with all new technologies, it still requires improvements. Different factors can affect CRISPR/Cas efficiency in zygotes, which influence the total cost and complexity for creating large-animal models for research. This study evaluated the importance of zygote cell cycle stage between early-injection (within 6 h post activation/fertilization) versus late-injection (14–16 h post activation/fertilization) when the CRISPR/Cas9 components were injected and the inhibition of the homologous recombination (HR) pathway of DNA repair on gene editing, embryo survival and development on embryos produced by fertilization, sperm injection, somatic cell nuclear transfer, and parthenogenetic activation technologies. Injections at the late cell cycle stage decreased embryo survival (measured as the proportion of unlysed embryos) and blastocyst formation (68.2%; 19.3%) compared to early-stage injection (86.3%; 28.8%). However, gene editing was higher in blastocysts from late-(73.8%) vs. early-(63.8%) injected zygotes. Inhibition of the HR repair pathway increased gene editing efficiency by 15.6% in blastocysts from early-injected zygotes without compromising embryo development. Our finding shows that injection at the early cell cycle stage along with HR inhibition improves both zygote viability and gene editing rate in pig blastocysts. Full article
(This article belongs to the Special Issue Cell Culture and Genome Modification Technologies: from Biology to Biomedical Applications)
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13 pages, 3565 KiB  
Article
Modulation of Epithelial Mesenchymal Transition after AGTR-1 Gene Edition by Crispr/Cas9 and Losartan Treatment in Mammary Tumor Cell Line: A Comparative Study between Human and Canine Species
by Marina Gobbe Moschetta-Pinheiro, Jucimara Colombo, Bianca Lara Venâncio de Godoy, Julia Ferreira Balan, Bianca Carlos Nascimento and Debora Aparecida Pires de Campos Zuccari
Life 2021, 11(12), 1427; https://doi.org/10.3390/life11121427 - 18 Dec 2021
Cited by 4 | Viewed by 2533
Abstract
Breast cancer is the most prevalent tumor type among women and female dogs. Tumor malignancy is characterized by the epithelial-to-mesenchymal transition (EMT) which leads to the metastasis formation. The inhibition of angiotensin II type I receptor (AGTR1) by an antagonist such as losartan [...] Read more.
Breast cancer is the most prevalent tumor type among women and female dogs. Tumor malignancy is characterized by the epithelial-to-mesenchymal transition (EMT) which leads to the metastasis formation. The inhibition of angiotensin II type I receptor (AGTR1) by an antagonist such as losartan can suppress angiogenesis, consequently contributing to the metastasis control. The aim of this study was to analyze the capacity of losartan and AGTR-1 gene edition to modulate the EMT process in triple negative/metastatic mammary tumor cells, compared to existing treatment protocols such as carboplatin. The cell lines CF41.Mg and MDA-MB-468, were cultured and treated with carboplatin, losartan, or submitted to AGTR-1 gene edition by CRISPR/Cas9. EMT markers and PARP-1 protein and gene expression were evaluated by immunofluorescence or immunocytochemistry and qRT-PCR, respectively. Cell migration capacity was also evaluated. For CF41.Mg and MDA-MB-468 cell lines, there was an increase in E-cadherin and a decrease in N-cadherin and PARP-1 protein and gene expression after treatment with carboplatin, losartan, both in combination and after AGTR-1 gene edition. There was a decrease in VEGF and PARP-1 protein and gene expression after AGTR-1 gene edition. Moreover, in both lines, reduction in invasion rate was observed after all treatments. Our data suggest that losartan and the gene edition of AGTR-1 by CRISPR/Cas9 were able to block the DNA repair and control the EMT process, such as carboplatin. The results in the canine species are unprecedented, as there are no data in the literature that demonstrate the action of losartan in this tumor type. Full article
(This article belongs to the Special Issue Cell Culture and Genome Modification Technologies: from Biology to Biomedical Applications)
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