Denilsa Pires Fernandes , João Vitor da Silva Viana , Luanna Lorenna Vieira Rodrigues , Lhara Ricarliany Medeiros de Oliveira , Sarah Leyenne Alves Sales , Maria Claudia dos Santos Luciano , Carlos Iberê Alves Freitas , Claudia Pessoa , Alexsandra Fernandes Pereira
{"title":"血清饥饿是诱导六带犰狳皮肤源性成纤维细胞G0/G1期细胞周期阻滞的最合适方法。","authors":"Denilsa Pires Fernandes , João Vitor da Silva Viana , Luanna Lorenna Vieira Rodrigues , Lhara Ricarliany Medeiros de Oliveira , Sarah Leyenne Alves Sales , Maria Claudia dos Santos Luciano , Carlos Iberê Alves Freitas , Claudia Pessoa , Alexsandra Fernandes Pereira","doi":"10.1016/j.tice.2024.102708","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><div>Several studies have evaluated different cell cycle synchronization methods to improve reprogramming efficiency aimed at wild species conservation. The six-banded armadillo is one of the wild mammals with significant ecological and biomedical interests but has not yet been evaluated for reprogramming purposes.</div></div><div><h3>Objective</h3><div>We investigated the effects in a time-dependent manner of serum starvation (SS; 0.5 % serum), contact inhibition (CI), and roscovitine (RSV 15 and 30 µM) on the cycle synchronization at G<sub>0</sub>/G<sub>1</sub> phase of six-banded armadillo fibroblasts by flow cytometric analysis. <em>Methods:</em> The morphology, viability, and apoptosis levels were determined by microscopic analysis.</div></div><div><h3>Results</h3><div>The results showed that SS significantly increased the percentage of cells arrested in the G<sub>0</sub>/G<sub>1</sub> phase in a time-dependent manner, where SS for 72 h (90.5 % ± 0.8) and 120 h (90.1 % ± 0.9) showed the highest value compared to SS for 24 h (86.1 % ± 0.9) and control group (69.7 % ± 0.8; <em>P<</em>0.05). However, neither CI nor RSV exposure significantly increased the arrest of fibroblasts at the G<sub>0</sub>/G<sub>1</sub> phase under any of the evaluated conditions (<em>P></em>0.05). In contrast to SS and RSV, CI significantly induced necrosis, as evidenced by an increase in dead cells (P < 0.05) over the evaluated periods, thereby disrupting the synchronization efforts. Moreover, all methods demonstrated similar morphology and cell viability rates over the evaluated times, as assessed by the trypan blue exclusion assay.</div></div><div><h3>Conclusion</h3><div>These results indicated that SS was the most suitable method for cell cycle modulation in G<sub>0</sub>/G<sub>1</sub> of six-banded armadillo fibroblasts, especially after 72–120 h of treatment. This finding has significant implications for future reprogramming efforts and conservation strategies for this unique species, as effective cell cycle synchronization can enhance the success of techniques, such as cloning and generation of induced cells to pluripotency.</div></div>","PeriodicalId":23201,"journal":{"name":"Tissue & cell","volume":"93 ","pages":"Article 102708"},"PeriodicalIF":2.7000,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Serum starvation as the most suitable method for inducing G0/G1 phase cell cycle arrest in six-banded armadillo (Euphractus sexcinctus) skin-derived fibroblast lines\",\"authors\":\"Denilsa Pires Fernandes , João Vitor da Silva Viana , Luanna Lorenna Vieira Rodrigues , Lhara Ricarliany Medeiros de Oliveira , Sarah Leyenne Alves Sales , Maria Claudia dos Santos Luciano , Carlos Iberê Alves Freitas , Claudia Pessoa , Alexsandra Fernandes Pereira\",\"doi\":\"10.1016/j.tice.2024.102708\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><div>Several studies have evaluated different cell cycle synchronization methods to improve reprogramming efficiency aimed at wild species conservation. The six-banded armadillo is one of the wild mammals with significant ecological and biomedical interests but has not yet been evaluated for reprogramming purposes.</div></div><div><h3>Objective</h3><div>We investigated the effects in a time-dependent manner of serum starvation (SS; 0.5 % serum), contact inhibition (CI), and roscovitine (RSV 15 and 30 µM) on the cycle synchronization at G<sub>0</sub>/G<sub>1</sub> phase of six-banded armadillo fibroblasts by flow cytometric analysis. <em>Methods:</em> The morphology, viability, and apoptosis levels were determined by microscopic analysis.</div></div><div><h3>Results</h3><div>The results showed that SS significantly increased the percentage of cells arrested in the G<sub>0</sub>/G<sub>1</sub> phase in a time-dependent manner, where SS for 72 h (90.5 % ± 0.8) and 120 h (90.1 % ± 0.9) showed the highest value compared to SS for 24 h (86.1 % ± 0.9) and control group (69.7 % ± 0.8; <em>P<</em>0.05). However, neither CI nor RSV exposure significantly increased the arrest of fibroblasts at the G<sub>0</sub>/G<sub>1</sub> phase under any of the evaluated conditions (<em>P></em>0.05). In contrast to SS and RSV, CI significantly induced necrosis, as evidenced by an increase in dead cells (P < 0.05) over the evaluated periods, thereby disrupting the synchronization efforts. Moreover, all methods demonstrated similar morphology and cell viability rates over the evaluated times, as assessed by the trypan blue exclusion assay.</div></div><div><h3>Conclusion</h3><div>These results indicated that SS was the most suitable method for cell cycle modulation in G<sub>0</sub>/G<sub>1</sub> of six-banded armadillo fibroblasts, especially after 72–120 h of treatment. This finding has significant implications for future reprogramming efforts and conservation strategies for this unique species, as effective cell cycle synchronization can enhance the success of techniques, such as cloning and generation of induced cells to pluripotency.</div></div>\",\"PeriodicalId\":23201,\"journal\":{\"name\":\"Tissue & cell\",\"volume\":\"93 \",\"pages\":\"Article 102708\"},\"PeriodicalIF\":2.7000,\"publicationDate\":\"2024-12-25\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Tissue & cell\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0040816624004099\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"ANATOMY & MORPHOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Tissue & cell","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0040816624004099","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"ANATOMY & MORPHOLOGY","Score":null,"Total":0}
Serum starvation as the most suitable method for inducing G0/G1 phase cell cycle arrest in six-banded armadillo (Euphractus sexcinctus) skin-derived fibroblast lines
Background
Several studies have evaluated different cell cycle synchronization methods to improve reprogramming efficiency aimed at wild species conservation. The six-banded armadillo is one of the wild mammals with significant ecological and biomedical interests but has not yet been evaluated for reprogramming purposes.
Objective
We investigated the effects in a time-dependent manner of serum starvation (SS; 0.5 % serum), contact inhibition (CI), and roscovitine (RSV 15 and 30 µM) on the cycle synchronization at G0/G1 phase of six-banded armadillo fibroblasts by flow cytometric analysis. Methods: The morphology, viability, and apoptosis levels were determined by microscopic analysis.
Results
The results showed that SS significantly increased the percentage of cells arrested in the G0/G1 phase in a time-dependent manner, where SS for 72 h (90.5 % ± 0.8) and 120 h (90.1 % ± 0.9) showed the highest value compared to SS for 24 h (86.1 % ± 0.9) and control group (69.7 % ± 0.8; P<0.05). However, neither CI nor RSV exposure significantly increased the arrest of fibroblasts at the G0/G1 phase under any of the evaluated conditions (P>0.05). In contrast to SS and RSV, CI significantly induced necrosis, as evidenced by an increase in dead cells (P < 0.05) over the evaluated periods, thereby disrupting the synchronization efforts. Moreover, all methods demonstrated similar morphology and cell viability rates over the evaluated times, as assessed by the trypan blue exclusion assay.
Conclusion
These results indicated that SS was the most suitable method for cell cycle modulation in G0/G1 of six-banded armadillo fibroblasts, especially after 72–120 h of treatment. This finding has significant implications for future reprogramming efforts and conservation strategies for this unique species, as effective cell cycle synchronization can enhance the success of techniques, such as cloning and generation of induced cells to pluripotency.
期刊介绍：
Tissue and Cell is devoted to original research on the organization of cells, subcellular and extracellular components at all levels, including the grouping and interrelations of cells in tissues and organs. The journal encourages submission of ultrastructural studies that provide novel insights into structure, function and physiology of cells and tissues, in health and disease. Bioengineering and stem cells studies focused on the description of morphological and/or histological data are also welcomed.
Studies investigating the effect of compounds and/or substances on structure of cells and tissues are generally outside the scope of this journal. For consideration, studies should contain a clear rationale on the use of (a) given substance(s), have a compelling morphological and structural focus and present novel incremental findings from previous literature.
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