{"title":"抗坏血酸对解冻后人脂肪干细胞形态的影响。","authors":"Komang Ardi Wahyuningsih, Karina Karina, Imam Rosadi, Iis Rosliana, Wismo Reja Subroto","doi":"10.21037/sci-2020-011","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Ascorbic acid-2-phosphate has been reported to play a role in cell division and to suppress aging of cell. However, post-thawed cell morphology on various concentration of ascorbic acid is still unclear. In this study, we aimed to observe the morphology of post-thawed adipose-derived stem cells (ADSCs) in medium containing L-ascorbic acid-2-phosphate (LAA2P) (50 and 100 µg/mL).</p><p><strong>Methods: </strong>The cells were isolated from adipose tissue. Isolated cells then cultured and cryopreserved in liquid nitrogen. We detected mRNA expression of type 1 collagen on day 5. Cell seeded in T25 flask using basal medium [Dulbecco's modified Eagle's medium (DMEM) only] as a control group, DMEM with 10% fetal bovine serum (FBS) and antibiotics as DMFA group, while DMFA with ascorbic acid (50 and 100 µg/mL) as ascorbic acid treatment group.</p><p><strong>Results: </strong>The results showed that the cells cultured in DMEM only attached until 96 hours of observation while serum groups with or without ascorbic acid supplementation showed the proliferation until 240 hours of observation. The highest spread size of cell was in a serum group without ascorbic acid supplementation and the highest yield of cells showed in a group with 50 µg/mL of ascorbic acid supplementation. Reduced mRNA expression of type 1 collagen which related to aging was showed in cells cultured without ascorbic acid supplementation.</p><p><strong>Conclusions: </strong>These results showed that ascorbic acid increased the cell division and suppressed the aging processes indicated by normal spread cell in size compared to cell cultured in DMFA without ascorbic acid supplementation.</p>","PeriodicalId":21938,"journal":{"name":"Stem cell investigation","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2020-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.21037/sci-2020-011","citationCount":"6","resultStr":"{\"title\":\"Effect of ascorbic acid on morphology of post-thawed human adipose-derived stem cells.\",\"authors\":\"Komang Ardi Wahyuningsih, Karina Karina, Imam Rosadi, Iis Rosliana, Wismo Reja Subroto\",\"doi\":\"10.21037/sci-2020-011\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Ascorbic acid-2-phosphate has been reported to play a role in cell division and to suppress aging of cell. However, post-thawed cell morphology on various concentration of ascorbic acid is still unclear. In this study, we aimed to observe the morphology of post-thawed adipose-derived stem cells (ADSCs) in medium containing L-ascorbic acid-2-phosphate (LAA2P) (50 and 100 µg/mL).</p><p><strong>Methods: </strong>The cells were isolated from adipose tissue. Isolated cells then cultured and cryopreserved in liquid nitrogen. We detected mRNA expression of type 1 collagen on day 5. Cell seeded in T25 flask using basal medium [Dulbecco's modified Eagle's medium (DMEM) only] as a control group, DMEM with 10% fetal bovine serum (FBS) and antibiotics as DMFA group, while DMFA with ascorbic acid (50 and 100 µg/mL) as ascorbic acid treatment group.</p><p><strong>Results: </strong>The results showed that the cells cultured in DMEM only attached until 96 hours of observation while serum groups with or without ascorbic acid supplementation showed the proliferation until 240 hours of observation. The highest spread size of cell was in a serum group without ascorbic acid supplementation and the highest yield of cells showed in a group with 50 µg/mL of ascorbic acid supplementation. Reduced mRNA expression of type 1 collagen which related to aging was showed in cells cultured without ascorbic acid supplementation.</p><p><strong>Conclusions: </strong>These results showed that ascorbic acid increased the cell division and suppressed the aging processes indicated by normal spread cell in size compared to cell cultured in DMFA without ascorbic acid supplementation.</p>\",\"PeriodicalId\":21938,\"journal\":{\"name\":\"Stem cell investigation\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2020-09-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.21037/sci-2020-011\",\"citationCount\":\"6\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Stem cell investigation\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.21037/sci-2020-011\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2020/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q1\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Stem cell investigation","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.21037/sci-2020-011","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2020/1/1 0:00:00","PubModel":"eCollection","JCR":"Q1","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 6
摘要
背景:抗坏血酸-2-磷酸已被报道在细胞分裂和抑制细胞衰老中起作用。然而,不同浓度抗坏血酸对解冻后细胞形态的影响尚不清楚。在这项研究中,我们旨在观察解冻后的脂肪来源干细胞(ADSCs)在含有l -抗坏血酸-2-磷酸(LAA2P)(50和100µg/mL)的培养基中的形态。方法:从脂肪组织中分离细胞。然后将分离的细胞培养并在液氮中冷冻保存。第5天检测1型胶原mRNA表达。细胞在T25烧瓶中接种,以基础培养基[Dulbecco's modified Eagle's medium (DMEM)]为对照组,DMEM中添加10%胎牛血清(FBS)和抗生素作为DMFA组,DMFA中添加抗坏血酸(50和100µg/mL)作为抗坏血酸处理组。结果:DMEM中培养的细胞仅粘附到96 h,而添加和不添加抗坏血酸的血清组细胞增殖到240 h。未添加抗坏血酸的血清组细胞扩散大小最大,添加50µg/mL抗坏血酸的血清组细胞产量最高。在不添加抗坏血酸的情况下,细胞中与衰老相关的1型胶原mRNA表达减少。结论:这些结果表明,与不添加抗坏血酸的DMFA培养的细胞相比,抗坏血酸增加了细胞分裂,抑制了正常细胞大小的衰老过程。
Effect of ascorbic acid on morphology of post-thawed human adipose-derived stem cells.
Background: Ascorbic acid-2-phosphate has been reported to play a role in cell division and to suppress aging of cell. However, post-thawed cell morphology on various concentration of ascorbic acid is still unclear. In this study, we aimed to observe the morphology of post-thawed adipose-derived stem cells (ADSCs) in medium containing L-ascorbic acid-2-phosphate (LAA2P) (50 and 100 µg/mL).
Methods: The cells were isolated from adipose tissue. Isolated cells then cultured and cryopreserved in liquid nitrogen. We detected mRNA expression of type 1 collagen on day 5. Cell seeded in T25 flask using basal medium [Dulbecco's modified Eagle's medium (DMEM) only] as a control group, DMEM with 10% fetal bovine serum (FBS) and antibiotics as DMFA group, while DMFA with ascorbic acid (50 and 100 µg/mL) as ascorbic acid treatment group.
Results: The results showed that the cells cultured in DMEM only attached until 96 hours of observation while serum groups with or without ascorbic acid supplementation showed the proliferation until 240 hours of observation. The highest spread size of cell was in a serum group without ascorbic acid supplementation and the highest yield of cells showed in a group with 50 µg/mL of ascorbic acid supplementation. Reduced mRNA expression of type 1 collagen which related to aging was showed in cells cultured without ascorbic acid supplementation.
Conclusions: These results showed that ascorbic acid increased the cell division and suppressed the aging processes indicated by normal spread cell in size compared to cell cultured in DMFA without ascorbic acid supplementation.
期刊介绍:
The Stem Cell Investigation (SCI; Stem Cell Investig; Online ISSN: 2313-0792) is a free access, peer-reviewed online journal covering basic, translational, and clinical research on all aspects of stem cells. It publishes original research articles and reviews on embryonic stem cells, induced pluripotent stem cells, adult tissue-specific stem/progenitor cells, cancer stem like cells, stem cell niche, stem cell technology, stem cell based drug discovery, and regenerative medicine. Stem Cell Investigation is indexed in PubMed/PMC since April, 2016.