In vitro cultivation of porcine limbal stem cells

Wharton’s jelly mesenchymal stem cells (WJ-MSCs) are multipotent stem cells that can be used in regenerative medicine. However, to reach the high therapeutic efficacy of WJ-MSCs, it is necessary to obtain a large amount of MSCs, which requires their extensive in vitro culturing. Numerous studies have shown that in vitro expansion of MSCs can lead to changes in cell behavior; cells lose their ability to proliferate, differentiate and migrate. One of the important measures of cells’ migration potential is their elasticity, determined by atomic force microscopy (AFM) and quantified by Young’s modulus. This work describes the elasticity of WJ-MSCs during in vitro cultivation. To identify the properties that enable transmigration, the deformability of WJ-MSCs that were able to migrate across the endothelial monolayer or Matrigel was analyzed by AFM. We showed that WJ-MSCs displayed differences in deformability during in vitro cultivation. This phenomenon seems to be strongly correlated with the organization of F-actin and reflects the changes characteristic for stem cell maturation. Furthermore, the results confirm the relationship between the deformability of WJ-MSCs and their migration potential and suggest the use of Young’s modulus as one of the measures of competency of MSCs with respect to their possible use in therapy.

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Lycopersicon esculentum lectin is a marker of transient amplifying cells in in vitro cultures of isolated limbal stem cells

Tissue and Cell ◽

10.1016/j.tice.2010.05.001 ◽

2010 ◽

Vol 42 (4) ◽

pp. 259-265 ◽

Cited By ~ 2

Author(s):

C. Vergallo ◽

T. Fonseca ◽

G. Pizzi ◽

L. Dini

Keyword(s):

Stem Cells ◽

Lycopersicon Esculentum ◽

In Vitro Cultures ◽

Limbal Stem Cells

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Purification and in vitro cultivation of archaeocytes (stem cells) of the marine sponge Hymeniacidon perleve (Demospongiae)

Cell and Tissue Research ◽

10.1007/s00441-006-0342-x ◽

2006 ◽

Vol 328 (1) ◽

pp. 223-237 ◽

Cited By ~ 24

Author(s):

Liming Sun ◽

Yuefan Song ◽

Yi Qu ◽

Xingju Yu ◽

Wei Zhang

Keyword(s):

Stem Cells ◽

Marine Sponge ◽

In Vitro Cultivation

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Effect of helium-neon laser on the limbal zone cells of human eye

Journal of New Medical Technologies eJournal ◽

10.12737/10419 ◽

2015 ◽

Vol 9 (1) ◽

pp. 0-0

Author(s):

Сабурова ◽

I. Saburova ◽

Копаева ◽

V. Kopaeva ◽

Копаев ◽

...

Keyword(s):

Stem Cells ◽

Cell Culture ◽

Progenitor Cells ◽

Low Energy ◽

Limbal Stem Cells ◽

Neon Laser ◽

Helium Neon Laser ◽

Stem And Progenitor Cells ◽

Helium Neon

Currently, a helium-neon laser is widely used in the treatment of degenerative, inflammatory and vascular diseases of the eye. It was found that the interaction of this radiation with different tissues, as a result of complex photochemical processes, is manifested by anti-inflammatory, desensitizing, resolving effects. Also, there is a stimulating effect on the processes of reparation and trophism. However, these observations relate to the use of laser for external irradiation, when the light goes through the air and the sclera or through the cornea. Research of the effect of endo-laser direct exposure of the fiber in the cavity of the eye, where light acts directly on epithelial or limbal cells, wasn’t carry out due to objective reasons (difficulty of conducting such research directly to patients). There is currently no data on the effect of helium-neon laser on the limbal area of the eye-ball, enriched endogenous stem and progenitor cells. They help repair damaged tissues in the eye (anterior and posterior segment) at various injuries. Cell culture, grown in vitro, are free from the influences of body systems, attract researchers as a unique model to study the behavior of cells in normal but also in response to external or internal factors. Currently, scientists have learned how to obtain and difficult to cultivate a culture of cells, in-cluding tissues of the human eye. They are genetically homogeneous population of cells growing in constant conditions. The viability and morphology of cells and their ultra-structure and various molecular biological cha-racteristics can be studied in cell culture, and to find the optimal conditions of laser irradiation, does not damage the cells. In the present work the authors studied the effect of low-energy helium-neon laser irradiation (632 nm) on limbal stem cells. Conducted in vitro studies have shown that the use of low-energy helium-neon laser irrad-iation (632 nm) has a positive effect on the monolayer culture of limbal stem cells. Absence of changes in cell phenotype and high proliferative activity indicate a stimulating effect of the investigated radiation on stem and progenitor cells, leading to the result of the activation of recovery and reduction of pathological changes at the cellular and organ and tissue levels.

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Corneal stromal mesenchymal stem cells: reconstructing a bioactive cornea and repairing the corneal limbus and stromal microenvironment

International Journal of Ophthalmology ◽

10.18240/ijo.2021.03.19 ◽

2021 ◽

Vol 14 (3) ◽

pp. 448-455

Author(s):

Xian-Ning Liu ◽

◽

Yun Chen ◽

Yao Wang ◽

◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Anterior Part ◽

Corneal Stroma ◽

Limbal Stem Cells ◽

Differentiation Potential ◽

Self Renewal ◽

Stromal Microenvironment ◽

Application Prospects

Corneal stroma-derived mesenchymal stem cells (CS-MSCs) are mainly distributed in the anterior part of the corneal stroma near the corneal limbal stem cells (LSCs). CS-MSCs are stem cells with self-renewal and multidirectional differentiation potential. A large amount of data confirmed that CS-MSCs can be induced to differentiate into functional keratocytes in vitro, which is the motive force for maintaining corneal transparency and producing a normal corneal stroma. CS-MSCs are also an important component of the limbal microenvironment. Furthermore, they are of great significance in the reconstruction of ocular surface tissue and tissue engineering for active biocornea construction. In this paper, the localization and biological characteristics of CS-MSCs, the use of CS-MSCs to reconstruct a tissue-engineered active biocornea, and the repair of the limbal and matrix microenvironment by CS-MSCs are reviewed, and their application prospects are discussed.

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Characterization of the secretory profile and exosomes of limbal stem cells in the canine species

PLoS ONE ◽

10.1371/journal.pone.0244327 ◽

2020 ◽

Vol 15 (12) ◽

pp. e0244327

Author(s):

Antonio J. Villatoro ◽

Cristina Alcoholado ◽

María del Carmen Martín-Astorga ◽

Gustavo Rico ◽

Viviana Fernández ◽

...

Keyword(s):

Stem Cells ◽

Ocular Surface ◽

Inhibitory Effect ◽

Proteomic Profile ◽

Limbal Stem Cells ◽

High Concentrations ◽

First Time

Limbal stem cells (LSCs) are a quiescent cell population responsible for the renewal of the corneal epithelium. Their deficiency is responsible for the conjunctivization of the cornea that is seen in different ocular pathologies, both in humans and in the canine species. The canine species represents an interesting preclinical animal model in ocular surface pathologies. However, the role of LSCs in physiological and pathological conditions in canine species is not well understood. Our objective was to characterize for the first time the soluble factors and the proteomic profile of the secretome and exosomes of canine LSCs (cLSCs). In addition, given the important role that fibroblasts play in the repair of the ocular surface, we evaluated the influence of the secretome and exosomes of cLSCs on their proliferation in vitro. Our results demonstrated a secretory profile of cLSCs with high concentrations of MCP-1, IL-8, VEGF-A, and IL-10, as well as significant production of exosomes. Regarding the proteomic profile, 646 total proteins in the secretome and 356 in exosomes were involved in different biological processes. Functionally, the cLSC secretome showed an inhibitory effect on the proliferation of fibroblasts in vitro, which the exosomes did not. These results open the door to new studies on the possible use of the cLSC secretome or some of its components to treat certain pathologies of the ocular surface in canine species.

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ФЕНОТИПОВІ ТА МОРФОЛОГІЧНІ ЗМІНИ КУЛЬТУРИ КЛІТИН КІСТКОВОГО МОЗКУ ЩУРІВ В ПРОЦЕСІ ЇХ КУЛЬТИВУВАННЯ

Scientific Messenger of LNU of Veterinary Medicine and Biotechnology ◽

10.15421/nvlvet6626 ◽

2016 ◽

Vol 18 (2(66)) ◽

pp. 126-132

Author(s):

A.I. Mazurkiewicz ◽

V.V. Kovpak ◽

O.S. Kovpak

Keyword(s):

Stem Cells ◽

Cell Culture ◽

Bone Marrow ◽

Bone Marrow Cells ◽

Morphological Changes ◽

Embryonic Stem ◽

In Vitro Cultivation ◽

Rat Bone ◽

Marrow Cells

Bone marrow is the only adult tissue which normally consists of immature undifferentiated and low differentiated cells which called stem cells and they are similar in structure to embryonic stem cells. But literature data analysis doesn't give an unambiguous answer regarding phenotypic and morphological changes of bone marrow cells culture of rats during their in vitro cultivation which necessitated further research.Investigate phenotypic and morphological changes of bone marrow cells culture of rats during their in vitro cultivation from first to fourth passage.We were used in these research bone marrow cells of rats from the first to the fourth passages. Microscopic analysis and evaluation morphological changes of bone marrow cells culture of rats during cultivation were carried out using inverted microscope Axiovert 40. Control of changes phenotype was performed by detecting CD markers (CD10, CD38, CD34, CD45, CD48, CD54, CD56, CD66e, CD96, CD227, CD326, pan–keratin). The evaluation was performed by the semi– quantitative method (H–Score).The research of primary culture of rat bone marrow cells showed that it morphologically heterogeneous, noted the small number of cells polygonal shape, surrounded by the fibroblast cells. During the cultivation cell culture becomes more homogenous at the expense of fibroblast–like cells. As a result of occurred the transition process from heterogeneous culture in zero passage to the most homogeneous culture in 4 passage. Immunophenotyping population of cell culture derived from rat bone marrow, revealed a high level of expression of pan–keratin; moderate level – CD34, CD48, CD66e, CD95; low level – CD38, CD45, CD56, CD227, CD326; lack of expression – CD10, CD54. Change of the expression of surface markers varies in each passage CD48, CD66e, CD95 increased significantly; CD38, SD45, SD326, pan–keratin reduced significantly. The markers CD34, CD 56, CD 227 were expressed on the one level from the first to the fourth passage. The expression of the CD10, CD54 markers during the study period was not identified.

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