The differentiation and transdifferentiation of epithelial cells in vitro – is it a new strategy in regenerative biomedicine?

Jankowski, Maurycy; Dyszkiewicz-Konwińska, Marta; Budna, Joanna; Knap, Sandra; Bryja, Artur; Borys, Sylwia; Kranc, Wiesława; Magas, Magdalena; Ješeta, Michal; Bukowska, Dorota; Antosik, Paweł; Brüssow, Klaus‐Peter; Machatkova, Marie; Bruska, M; Nowicki, Michał; Zabel, Maciej; Kempisty, Bartosz

doi:10.2478/acb-2018-0005

Cited by 8 publications

(6 citation statements)

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“…It is well known that the advancement of modern medicine will be highly dependent on the development of knowledge regarding stem cells [1][2][3][4][5][6]. While some studies focus on embryonic or induced pluripotent stem cells, the limited knowledge of stem cell regulatory mechanisms, as well as the significant risks associated with their administration, effectively delays any therapeutic approaches based on these cell types [7,8].…”

Section: Introductionmentioning

confidence: 99%

Expression Profile of New Marker Genes Involved in Differentiation of Canine Adipose-Derived Stem Cells into Osteoblasts

Jankowski

Kaczmarek

Wąsiatycz

et al. 2021

IJMS

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Next-generation sequencing (RNAseq) analysis of gene expression changes during the long-term in vitro culture and osteogenic differentiation of ASCs remains to be important, as the analysis provides important clues toward employing stem cells as a therapeutic intervention. In this study, the cells were isolated from adipose tissue obtained during routine surgical procedures and subjected to 14-day in vitro culture and differentiation. The mRNA transcript levels were evaluated using the Illumina platform, resulting in the detection of 19,856 gene transcripts. The most differentially expressed genes (fold change >|2|, adjusted p value < 0.05), between day 1, day 14 and differentiated cell cultures were extracted and subjected to bioinformatical analysis based on the R programming language. The results of this study provide molecular insight into the processes that occur during long-term in vitro culture and osteogenic differentiation of ASCs, allowing the re-evaluation of the roles of some genes in MSC progression towards a range of lineages. The results improve the knowledge of the molecular mechanisms associated with long-term in vitro culture and differentiation of ASCs, as well as providing a point of reference for potential in vivo and clinical studies regarding these cells’ application in regenerative medicine.

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Section: Introductionmentioning

confidence: 99%

Expression Profile of New Marker Genes Involved in Differentiation of Canine Adipose-Derived Stem Cells into Osteoblasts

Jankowski

Kaczmarek

Wąsiatycz

et al. 2021

IJMS

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“…e cells were cultured at 38°C in a humidified atmosphere of 5% CO 2 . e culture medium was changed every 3 days [13,14].…”

Section: Endometrial Cell (Ec) Selection and Culturementioning

confidence: 99%

“…FITC was excited at 488 nm with an argon laser, with emissions being imaged through a 505-530 nm filter. All confocal microscopic images were analyzed using Imaris 7.2 software (BitPlane, Zurich, Switzerland) [13,14]. In order to validate the specificity of antibodies for all samples, we used control experiment where only secondary antibody was used, and to analyze the changes in the expression of proteins, we used 10 images for each sample.…”

Section: Confocal Microscopy Analysis Of Connexin Expression and Distmentioning

confidence: 99%

Expression of Selected Connexin and Aquaporin Genes and Real-Time Proliferation of Porcine Endometrial Luminal Epithelial Cells in Primary Culture Model

Wojtanowicz-Markiewicz

Kulus

Knap

et al. 2020

BioMed Research International

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Luminal epithelial cells are the first embryonic-maternal contact site undergoing very specific changes associated with reproductive processes. Cells prepare for embryo development by increasing their volume, with the help of aquaporins that provide a transcellular path of rapid water movement during the secretion and absorption of fluids, as well as connexins enabling the flow of inorganic ions and small molecules. In this work, we have examined how AQPs and Cx’s behave in luminal epithelium primary cell culture. Cells obtained from porcine specimen during slaughter were primarily in vitro cultured for 7 days. Their proliferation patterns were then analyzed using RTCA, with the expression of genes of interest evaluated with the use of immunofluorescence and RT-qPCR. The results of these changes of gene of interest expression were analyzed on each of the seven days of the porcine luminal primary cell culture. Our study showed that the significant changes were noted in the case of Cx43, whose level of protein expression and distribution increases after 120 hours of culture, when the cells enter the lag phase, and maintains an upward trend until the end of the culture. We noted an increase in AQP4, AQP7, AQP8, and AQP11 levels throughout the entire culture period, while the largest differences in expression were found in AQP3, AQP4, and AQP10. The obtained results could become a point of reference for further in vivo and clinical research. Experiments conducted with these proteins showed that they influence the endometrial fluid content during the oestrous cycle and participate in the process of angiogenesis, which intensifies during endometrial development.

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“…These experiments indicated a huge stem-like specificity and plasticity of human GCs in primary culture. These results label GCs as potential candidates for future use as allografts in regenerative and reconstructive medicine (Jankowski et al 2018; Kranc et al 2018). In this study, we aim to recognize the expression patterns of genes involved in their in vitro proliferation and differentiation.…”

Section: Introductionmentioning

confidence: 98%

Genes responsible for proliferation, differentiation, and junction adhesion are significantly up-regulated in human ovarian granulosa cells during a long-term primary in vitro culture

Kranc

Brązert

Budna

et al. 2018

Histochem Cell Biol

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The human ovarian granulosa cells (GCs) surround the oocyte and form the proper architecture of the ovarian follicle. The ability of GCs to proliferate and differentiate in the conditions of in vitro culture has been proven. However, there is still a large field for extensive investigation of molecular basics, as well as marker genes, responsible for these processes. This study aimed to find the new marker genes, encoding proteins that regulate human GCs in vitro capability for proliferation and differentiation during long-term primary culture. The human follicular GCs were collected from hyper-stimulated ovarian follicles during IVF procedures and transferred to a long-term in vitro culture. The culture lasted for 30 days, with RNA samples isolated at days 1, 7, 15, 30. Transcriptomic analysis was then performed with the use of Affymetrix microarray. Obtained results were then subjected to bioinformatical evaluation and sorting. After subjecting the datasets to KEGG analysis, three differentially expressed ontology groups “cell differentiation” (GO:0030154), “cell proliferation” (GO:0008283) and “cell–cell junction organization” (GO:0045216) were chosen for further investigation. All three of those ontology groups are involved in human GCs’ in vitro lifespan, proliferation potential, and survival capability. Changes in expression of genes of interest belonging to the chosen GOs were validated with the use of RT-qPCR. In this manuscript, we suggest that VCL, PARVA, FZD2, NCS1 , and COL5A1 may be recognized as new markers of GC in vitro differentiation, while KAT2B may be a new marker of their proliferation. Additionally, SKI, GLI2, FERMT2 , and CDH2 could also be involved in GC in vitro proliferation and differentiation processes. We demonstrated that, in long-term in vitro culture, GCs exhibit markers that suggest their ability to differentiate into different cells types. Therefore, the higher expression profile of these genes may also be associated with the induction of cellular differentiation processes that take place beyond the long-term primary in vitro culture. Electronic supplementary material The online version of this article (10.1007/s00418-018-1750-1) contains supplementary material, which is available to authorized users.

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The differentiation and transdifferentiation of epithelial cells in vitro – is it a new strategy in regenerative biomedicine?

Cited by 8 publications

References 50 publications

Expression Profile of New Marker Genes Involved in Differentiation of Canine Adipose-Derived Stem Cells into Osteoblasts

Expression Profile of New Marker Genes Involved in Differentiation of Canine Adipose-Derived Stem Cells into Osteoblasts

Expression of Selected Connexin and Aquaporin Genes and Real-Time Proliferation of Porcine Endometrial Luminal Epithelial Cells in Primary Culture Model

Genes responsible for proliferation, differentiation, and junction adhesion are significantly up-regulated in human ovarian granulosa cells during a long-term primary in vitro culture

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