Approaches to the use of stem cells in regenerative medicine

Yessentayeva, Suriya Ye; Orakbay, Lyazat Zh; Adilhanova, Azhar; Yessimov, Nabi

doi:10.1016/j.ab.2022.114608

Cited by 7 publications

(7 citation statements)

References 15 publications

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“…In the present study, a monolayer culturing technique was used to compare the effects of a novel culture medium on the OKF6/TERT-2 cell line illustrating that DFK is a viable alternative medium, and more importantly, showing that this oral epithelial cell line remains an appropriate model of oral physiology during multiple conditions. Recently, organotypic 3-dimensional cultures with multiple cell layers have been developed to reflect the oral environment more accurately and therefore increase the pertinence of in vitro experimentation [88][89][90][91], which is a limitation of this investigation. This study was conducted on the premise that a novel culture medium should be evaluated on a monolayer before extension to 3D models.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Culture Media for in-vitro Expansion of Oral Epithelial Keratinocytes: Implications for Testing E-liquid Flavors

Cuadra¹,

Shamim²,

Shah³

et al. 2023

Preprint

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Background: Understanding in vitro expansion of OKF6/TERT-2 oral epithelial cells is important for studying molecular biology of disease and pathology affecting the oral cavity. The media used for any cell culture is paramount in terms of efficient output. Therefore, this study aims to compare two different media for OKF6/TERT-2 cultures: Keratinocyte Serum-Free Medium (KSFM) and a composite medium comprised of DMEM/F-12 mixed with KSFM (referred to as DFK). For application purposes, this investigation also compares the toxicological effects of flavored electronic cigarette liquids (E-liquids) on OKF6/TERT-2 cultures grown in both media. Methods: Cells were grown in KSFM and DFK media and cellular growth, morphology, gene expression of mucins and tight junctions, as well as wound-healing were determined. Additionally, cellular viability and cytotoxicity were indexed after E-liquid exposures. Results: While overall cellular morphologies remained unaltered, cells grown in DFK reached confluency faster. Except for claudin-1, there is no appreciable difference in expression of the other genes tested. Additionally, cultures in DFK appear more sensitive to E-liquids ± flavors. Conclusions: DFK is an alternative medium for cultivation of OKF6/TERT-2 cells to study molecular biology of disease and pathology, such as their responses to E-liquids ± flavors.

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Section: Discussionmentioning

confidence: 99%

Comparison of Culture Media for in-vitro Expansion of Oral Epithelial Keratinocytes: Implications for Testing E-liquid Flavors

Cuadra¹,

Shamim²,

Shah³

et al. 2023

Preprint

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“…For example, this study involved the use of an existing DPSC biorepository, which may have influenced the outcomes of the study due to the long-term effects associated with cryopreservation and storage [ 60 , 61 ]. In addition, this study was only able to evaluate DPSCs from a single repository; therefore, these results should be confirmed using other DPSC biorepositories and ex vivo samples [ 62 ]. Finally, other factors, such as the expression of microRNAs, may be important factors to consider in future studies of DPSC responsiveness to the ECM as in other recent studies of MSCs [ 63 , 64 , 65 ].…”

Section: Discussionmentioning

confidence: 99%

Differential Effects of Extracellular Matrix Glycoproteins Fibronectin and Laminin-5 on Dental Pulp Stem Cell Phenotypes and Responsiveness

Lee

Bae

Kim

et al. 2023

JFB

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Dental pulp stem cells (DPSCs) are mesenchymal stem cells (MSCs) with the potential to differentiate in a limited number of other tissue types. Some evidence has suggested the modulation of DPSC growth may be mediated, in part, by exogenous extracellular matrix (ECM) glycoproteins, including fibronectin (FN) and laminin-5 (LN5). Although preliminary research suggests that some ECM glycoproteins may work as functional biomaterials to modulate DPSC growth responses, the primary goal of this project is to determine the specific effects of FN and LN5 on DPSC growth and viability. Using an existing DPSC repository, n = 16 DPSC isolates were cultured and 96-well growth assays were performed, which revealed FN, LN5 and the combination of these were sufficient to induce statistically significant changes in growth among five (n = 5) DPSC isolates. In addition, the administration of FN (either alone or in combination) was sufficient to induce the expression of alkaline phosphatase (ALP) and dentin sialophosphoprotein (DSPP), while LN5 induced the expression of ALP only, suggesting differential responsiveness among DPSCs. Moreover, these responses appeared to correlate with the expression of MSC biomarkers NANOG, Oct4 and Sox2. These results add to the growing body of evidence suggesting that functional biomaterials, such as ECM glycoproteins FN and LN5, are sufficient to induce phenotypic and differentiation-specific effects in a specific subset of DPSC isolates. More research will be needed to determine which biomarkers or additional factors are necessary and sufficient to induce the differentiation and development of DPSCs ex vivo and in vitro for biomedical applications.

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“…Since our research interests focus on the use of electronic cigarettes and the effects of these on oral mucosa, this study compared the applicability of DFK or KSFM on OKF6/TERT-2 cultures exposed to E-liquids ± flavors. An oral epithelial cell line, amenable to multiple culture conditions, without compromising the phenotype of the model, will not only facilitate further research in these areas using monolayer culturing techniques but also provide a basis for further improved organotypic 3D cultures, a more realistic representation of in vivo tissues [91][92][93][94].…”

Section: Discussionmentioning

confidence: 99%

“…While our results are novel and contribute to the field of in vitro oral biology, they are not without limitations. For example, organotypic 3-dimensional cultures with multiple cell layers reflect the oral environment more accurately and therefore increase the pertinence of in vitro experimentation [91][92][93][94]. However, the present study was conducted on the premise that a novel culture medium should be evaluated on a monolayer before extension to more complex 3D models.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Culture Media for in-vitro Expansion of Oral Epithelial Keratinocytes: Implications for Testing E-liquid Flavors

Cuadra¹,

Shamim²,

Shah³

et al. 2023

Preprint

View full text Add to dashboard Cite

Background: Expansion of OKF6/TERT-2 oral epithelial cells in vitro is important for studying the molecular biology of disease and pathology affecting the oral cavity. Keratinocyte Serum-Free Medium (KSFM) is the medium of choice for this cell line. This study compares three media for OKF6/TERT-2 cultures: KSFM, Dulbecco’s Modified Eagle Medium/Nutrient Mixture of Hams F-12 (DMEM/F12) and a composite medium comprised of DMEM/F-12 and KSFM (1:1 v/v), referred as DFK. The toxicological effects of electronic cigarette liquids (E-liquids) on OKF6/TERT-2 cells cultured in these media were also compared. Methods: Cells were cultured in KSFM, DMEM/F12 or DFK and cellular morphology, growth, wound healing and gene expression of mucins and tight junctions were evaluated. Additionally, cytotoxicity was determined after E-liquid exposures. Results: Switching from KSFM to DMEM/F12 or DFK 24-hours post-seeding leads to typical cellular morphologies, and these cultures reach confluency faster than those in KSFM. Wound-healing recovery occurred fastest in DFK. Except for claudin-1, there is no difference in expression of the other genes tested. Additionally, E-liquid cytotoxicity appears to be amplified in DFK cultures. Conclusions: DMEM/F12 and DFK are alternative media for OKF6/TERT-2 cell culture to study molecular biology of disease and pathology, provided cells are initially seeded in KSFM.

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Approaches to the use of stem cells in regenerative medicine

Cited by 7 publications

References 15 publications

Comparison of Culture Media for <em>in-vitro</em> Expansion of Oral Epithelial Keratinocytes: Implications for Testing E-liquid Flavors

Comparison of Culture Media for <em>in-vitro</em> Expansion of Oral Epithelial Keratinocytes: Implications for Testing E-liquid Flavors

Differential Effects of Extracellular Matrix Glycoproteins Fibronectin and Laminin-5 on Dental Pulp Stem Cell Phenotypes and Responsiveness

Comparison of Culture Media for <em>in-vitro</em> Expansion of Oral Epithelial Keratinocytes: Implications for Testing E-liquid Flavors

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