Behavioral Changes of Multipotent Mesenchymal Stromal Cells in Contact with Synthetic Calcium Phosphates in vitro

Litvinova, L. S.; Шуплецова, В. В.; Хазиахматова, О. Г.; Yurova, K. A.; Малащенко, В. В.; Melashchenko, E. S.; Тодосенко, Н. М.; Khlusova, M. Yu.; Sharkeev, Yu. P.; Комарова, Е. Г.; Sedelnikova, M. B.; Shunkin, E. O.; Хлусов, И. А.

doi:10.1134/s1990519x18020062

Cited by 13 publications

(15 citation statements)

References 20 publications

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“…More than 95% of fibroblast-like or spindle adherent cells (Fig. 1) positively stained with al 123 (osteoblasts), alcian blue (chondrocytes), and oil red (adipocytes) expressed CD73, CD90, and CD105 124 did not display CD45, CD34, CD20, and CD14 markers (less 5%) (Litvinova et al, 2018). These find 125 their correspondence to the morphological criteria of MMSCs (Dominici et al, 2006;Bourin et al, 2013 126 127 128…”

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confidence: 67%

“…To determine the stem nature of human adipose-derived MMSCs (hAMMSCs), they were cultivated in the medium with a StemPro® Differentiation Kit (Thermo Fisher Scientific, United States) for 21 days with medium exchange every 3-4 days. As we published previously (Litvinova et al, 2018), chondrogenic, osteogenic and adipogenic differentiations were detected by cell positive staining with alcian blue (Sigma-Aldrich, United States) and visualized proteoglycan synthesis by chondrocytes; alizarin red S (Sigma-Aldrich, United States) identified mineralization of intercellular substances in osteoblasts, and oil red (Sigma-Aldrich, United States) detected neutral triglycerides and lipids in adipocytes. This confirmed that obtained cells belonged to the pool of stem cells according to the minimal morphological criteria of the International Society for Cellular Therapy (Dominici et al, 2006).…”

Section: Mmscs Isolationmentioning

confidence: 95%

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The possibilities of Cell-IQ phase-contrast microscopy for a continuous real-time observation of multipotent mesenchymal stromal cell culture

Melashchenko

Шуплецова

Хазиахматова

et al. 2018

EREM

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Continuous monitoring of multipotent mesenchymal stromal cells (MMSCs) is a promising tool that could be used in cellular biology, environmental research and biotechnology to study in vitro real-time morphology and behavioural response of stem cells alone, as well at contacts with other cells and for controlling a sustainable production of scaffolds for tissue engineering. The in vitro processes of human adipose-derived MMSCs (hAMMSCs) morphology, motility, cell division, and secretion were studied by means of Cell-IQ v2 MLF (CM Technologies Oy, Finland) integrated phase-contrast microscopic platform for a continuous real-time Environmental Research, Engineering and Management 2018/74/3 surveillance imaging of living cells. 70 μL suspension (5×10 4 viable karyocytes) of the cells was applied into the centre of the wells of 12-well plastic plates, and cells were allowed to adhere in a moist chamber for 120 min. Non-adherent cells were washed, and the wells were carefully filled with 1.5 mL of a nutrient medium DMEM/ F12 (1 : 1) without osteogenic additions. Cells were cultured for 14 days at 100% humidity in a 5% carbon dioxide atmosphere at 37°C until a monolayer formation. Digital images of cell culture growth were captured every 45 min. The cells were positively stained with alizarin red (osteoblasts), alcian blue (chondrocytes), or oil red (adipocytes). More than 95% of attached cells expressed CD73, CD90, and CD105 markers, mainly. Thus, the cells corresponded to the morphological criteria of MMSCs. The Cell-IQ system allowed establishing 182 μm/h linear velocity of free (until the cell contacts) motility of spindle or fibroblast-like cells. Maximum number of cells achieved 136 cells per field of view; 13-24% of cells divided each 1-3 h until a monolayer was formed. Chemokine cooperation between hAMMSCs and poor macrophages intermixture was proposed. Cell-IQ could be useful for in vitro real-time imaging of cell subpopulations and/or their response to biodegradable scaffolds and/or (micro)environmental factors.

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confidence: 67%

Section: Mmscs Isolationmentioning

confidence: 95%

The possibilities of Cell-IQ phase-contrast microscopy for a continuous real-time observation of multipotent mesenchymal stromal cell culture

Melashchenko

Шуплецова

Хазиахматова

et al. 2018

EREM

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“…These ions can regulate the expression of MSC markers [ 30 ]. Indeed, contact of hAMSCs with CaP-coated samples for 21 days was found to suppress the immunophenotypic traits of hAMSCs adhered to plastic as the relief of the CaP coating increased, while the population of hematopoietic cells (CD45 + CD34 + CD14 + CD20 + ) increased [ 22 ].…”

Section: Resultsmentioning

confidence: 99%

“…Microarc oxidation (MAO) or plasma electrolytic oxidation (PEO) is one the appropriate method to deposit CaP coatings for biomedical applications [ 18 , 19 , 20 ]. We found previously ECM mineralization of 21-day MSC culture both on rough surface [ 21 ] and on plastic around the microarc CaP-coated titanium (Ti) samples [ 22 ]. ALP and OCN expression [ 21 ] and secretion [ 23 ] accelerated by in vitro contact with microarc CaP coating.…”

Section: Introductionmentioning

confidence: 99%

Gene Expression Regulation and Secretory Activity of Mesenchymal Stem Cells upon In Vitro Contact with Microarc Calcium Phosphate Coating

Litvinova

Yurova

Шуплецова

et al. 2020

IJMS

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The manufacture of biomaterial surfaces with desired physical and chemical properties that can directly induce osteogenic differentiation without the need for biochemical additives is an excellent strategy for controlling the behavior of mesenchymal stem cells (MSCs) in vivo. We studied the cellular and molecular reactions of MSCs to samples with a double-sided calcium phosphate (CaP) coating and an average roughness index (Ra) of 2.4–4.6 µm. The study aimed to evaluate the effect of a three-dimensional matrix on the relative mRNA expression levels of genes associated with the differentiation and maturation of MSCs toward osteogenesis (RUNX2, BMP2, BMP6, BGLAP, and ALPL) under conditions of distant interaction in vitro. Correlations were revealed between the mRNA expression of some osteogenic and cytokine/chemokine genes and the secretion of cytokines and chemokines that may potentiate the differentiation of cells into osteoblasts, which indicates the formation of humoral components of the extracellular matrix and the creation of conditions supporting the establishment of hematopoietic niches.

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“…The experiment was performed according to previously described methods [ 28 , 29 ] with some modifications. All experiments were performed in laminar sterile air.…”

Section: Methodsmentioning

confidence: 99%

Costimulatory Effect of Rough Calcium Phosphate Coating and Blood Mononuclear Cells on Adipose-Derived Mesenchymal Stem Cells In Vitro as a Model of In Vivo Tissue Repair

et al. 2020

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Calcium phosphate (CaP) materials do not always induce ectopic vascularization and bone formation; the reasons remain unclear, and there are active discussions of potential roles for post-implantation hematoma, circulating immune and stem cells, and pericytes, but studies on adipose-derived stem cells (AMSCs) in this context are lacking. The rough (average surface roughness Ra = 2–5 µm) scaffold-like CaP coating deposited on pure titanium plates by the microarc oxidation method was used to investigate its subcutaneous vascularization in CBA/CaLac mice and in vitro effect on cellular and molecular crosstalk between human blood mononuclear cells (hBMNCs) and AMSCs (hAMSCs). Postoperative hematoma development on the CaP surface lasting 1–3 weeks may play a key role in the microvessel elongation and invasion into the CaP relief at the end of the 3rd week of injury and BMNC migration required for enhanced wound healing in mice. Satisfactory osteogenic and chondrogenic differentiation but poor adipogenic differentiation of hAMSCs on the rough CaP surface were detected in vitro by differential cell staining. The fractions of CD73+ (62%), CD90+ (0.24%), and CD105+ (0.41%) BMNCs may be a source of autologous circulating stem/progenitor cells for the subcutis reparation, but allogenic hBMNC participation is mainly related to the effects of CD4+ T cells co-stimulated with CaP coating on the in vitro recruitment of hAMSCs, their secretion of angiogenic and osteomodulatory molecules, and the increase in osteogenic features within the period of in vivo vascularization. Cellular and molecular crosstalk between BMNCs and AMSCs is a model of effective subcutis repair. Rough CaP surface enhanced angio- and osteogenic signaling between cells. We believe that preconditioning and/or co-transplantation of hAMSCs with hBMNCs may broaden their potential in applications related to post-implantation tissue repair and bone bioengineering caused by microarc CaP coating.

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Behavioral Changes of Multipotent Mesenchymal Stromal Cells in Contact with Synthetic Calcium Phosphates in vitro

Cited by 13 publications

References 20 publications

The possibilities of Cell-IQ phase-contrast microscopy for a continuous real-time observation of multipotent mesenchymal stromal cell culture

The possibilities of Cell-IQ phase-contrast microscopy for a continuous real-time observation of multipotent mesenchymal stromal cell culture

Gene Expression Regulation and Secretory Activity of Mesenchymal Stem Cells upon In Vitro Contact with Microarc Calcium Phosphate Coating

Costimulatory Effect of Rough Calcium Phosphate Coating and Blood Mononuclear Cells on Adipose-Derived Mesenchymal Stem Cells In Vitro as a Model of In Vivo Tissue Repair

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