Enhancing Proliferation of Stem Cells from Human Exfoliated Deciduous Teeth (SHED) through hTERT Expression while Preserving Stemness and Multipotency.

IF 4.5 3区医学 Q2 CELL & TISSUE ENGINEERING

Stem Cell Reviews and Reports Pub Date : 2024-10-01 Epub Date: 2024-06-15 DOI:10.1007/s12015-024-10746-y

Pooja Yadav, Ravina Vats, Sapna Wadhwa, Afsareen Bano, Ritu Namdev, Monika Gupta, Rashmi Bhardwaj

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Abstract

Background: Stem cells from human exfoliated deciduous teeth (SHED) hold promise in regenerative medicine owing to their multipotent capabilities resembling mesenchymal stem cells (MSCs). Despite their potential, SHED have not been extensively investigated because their limited lifespan and unavailability of cell-lines pose challenges for therapeutic applications. This study investigated the effect of ectopic human telomerase reverse transcriptase (hTERT) expression on SHEDs' proliferation while preserving stemness and genomic integrity.

Methods: Deciduous teeth were collected from children aged 6-10 years. After isolation and characterization, the SHED were transduced with pBabe-puro-hTERT retrovirus to establish SHED cell-line, which was evaluated and compared with pBabe-puro (mock control) for stemness, multipotency and growth attributes through flow cytometry, trilineage differentiation, and growth kinetics. We also estimated hTERT gene expression, genomic integrity, and validated cell-line through STR analysis.

Results: Following hTERT transduction, SHED displayed elevated hTERT gene expression while retaining fibroblast-like morphology and mesenchymal stem cell markers. Moreover, after hTERT transduction cellular shape remained same along with increased replicative lifespan and proliferation potential. SHED-hTERT cells exhibited multi-potency and maintained stemness, as evidenced by surface marker expression and multilineage differentiation. Furthermore, genomic integrity was not affected by hTERT integration, as confirmed by STR analysis and CDKN2A gene assessment.

Conclusion: Ectopic hTERT expression in SHED successfully prolonged their replicative lifespan and improved their ability to proliferate and migrate, while preserving their stemness, multipotency and genomic integrity, suggesting minimal carcinogenic risk. Establishment of SHED cell-line holds potential in regenerative medicine applications, especially in cell-based drugs and tissue engineering experiments.

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通过 hTERT 表达增强人类脱落牙齿（SHED）干细胞的增殖，同时保留干性和多潜能性。

背景：从人类脱落牙齿（SHED）中提取的干细胞具有类似间充质干细胞（MSCs）的多能性，因此有望用于再生医学。尽管SHED具有潜力，但由于其寿命有限且无法获得细胞系，因此尚未对其进行广泛研究，这给治疗应用带来了挑战。本研究调查了异位人类端粒酶逆转录酶（hTERT）表达对SHEDs增殖的影响，同时保留了干性和基因组完整性：方法：收集6-10岁儿童的乳牙。通过流式细胞术、三系分化和生长动力学，对SHED细胞系的干性、多能性和生长属性进行评估，并与pBabe-puro（模拟对照）进行比较。我们还估算了 hTERT 基因表达、基因组完整性，并通过 STR 分析验证了细胞系：结果：hTERT转导后，SHED的hTERT基因表达升高，同时保留了成纤维细胞样形态和间充质干细胞标记。此外，经 hTERT 转导后，细胞形态保持不变，复制寿命和增殖潜力也有所增加。SHED-hTERT细胞表现出多种潜能并保持干性，这一点可以从表面标记表达和多线分化得到证明。此外，基因组完整性不受 hTERT 整合的影响，STR 分析和 CDKN2A 基因评估证实了这一点：结论：在SHED中异位表达hTERT成功地延长了它们的复制寿命，提高了它们的增殖和迁移能力，同时保留了它们的干性、多能性和基因组完整性，表明致癌风险极低。SHED细胞系的建立具有再生医学应用的潜力，特别是在基于细胞的药物和组织工程实验中。

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来源期刊

Stem Cell Reviews and Reports 医学-细胞生物学

CiteScore

9.30

自引率

4.20%

发文量

审稿时长

3 months

期刊介绍： The purpose of Stem Cell Reviews and Reports is to cover contemporary and emerging areas in stem cell research and regenerative medicine. The journal will consider for publication: i) solicited or unsolicited reviews of topical areas of stem cell biology that highlight, critique and synthesize recent important findings in the field. ii) full length and short reports presenting original experimental work. iii) translational stem cell studies describing results of clinical trials using stem cells as therapeutics. iv) papers focused on diseases of stem cells. v) hypothesis and commentary articles as opinion-based pieces in which authors can propose a new theory, interpretation of a controversial area in stem cell biology, or a stem cell biology question or paradigm. These articles contain more speculation than reviews, but they should be based on solid rationale. vi) protocols as peer-reviewed procedures that provide step-by-step descriptions, outlined in sufficient detail, so that both experts and novices can apply them to their own research. vii) letters to the editor and correspondence. In order to facilitate this exchange of scientific information and exciting novel ideas, the journal has created five thematic sections, focusing on: i) the role of adult stem cells in tissue regeneration; ii) progress in research on induced pluripotent stem cells, embryonic stem cells and mechanism governing embryogenesis and tissue development; iii) the role of microenvironment and extracellular microvesicles in directing the fate of stem cells; iv) mechanisms of stem cell trafficking, stem cell mobilization and homing with special emphasis on hematopoiesis; v) the role of stem cells in aging processes and cancerogenesis.