Endoboost plus is the medium of choice for the cell mass expansion of ecfc isolated from adult blood

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Abstract

Endothelial colony-forming cells (ECFC) have a powerful clonogenic and reparative potential, which makes them a promising material for cell therapy, experimental biology and medicine. The ability to rapidly expand the cell mass is the key to the use of ECFCs in these areas. We are developing the composition of nutrient medium for EndoBoost and EndoBoost Plus endothelial cells. Endothelial cell growth medium2 (EGM2) is recognised as the ‘gold standard’ in ECFC cultivation. The aim of our study was to comparatively evaluate the efficacy of EGM2, EndoBoost and EndoBoost Plus nutrient media for ECFC culture growth. Maximum proliferative activity of ECFC was detected in EndoBoost Plus medium, EndoBoost was found to be less active and the lowest result corresponded to EGM2. Thus, EndoBoost Plus is the preferred medium for cell culture of ECFCs isolated from adult peripheral blood.

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About the authors

V. G. Matveeva

Research Institute for Complex Issues of Cardiovascular Diseases

Author for correspondence.
Email: matveeva_vg@mail.ru
Russian Federation, 650002, Kemerovo

D. K. Shishkova

Research Institute for Complex Issues of Cardiovascular Diseases

Email: matveeva_vg@mail.ru
Russian Federation, 650002, Kemerovo

Е. А. Torgunakova

Research Institute for Complex Issues of Cardiovascular Diseases

Email: matveeva_vg@mail.ru
Russian Federation, 650002, Kemerovo

A. G. Kutikhin

Research Institute for Complex Issues of Cardiovascular Diseases

Email: matveeva_vg@mail.ru
Russian Federation, 650002, Kemerovo

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2. Fig. 1. Typical histograms of antigen fluorescence on the cell membrane obtained using flow cytometry (fluorescently labeled antibodies - red curves; isotype control - black curves): a - endothelial antigens CD31, CD146, CD133, CD105; b - CD34; c - hematopoietic linear marker CD45 and CD14; d - myeloid CD90.

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3. Fig. 2. Typical morphology of ECFC colonies (a), as well as fluorescence of surface (CD31, CD144) and intracellular (VWF, F-actin) markers and extracellular matrix proteins synthesized by cells (fibronectin, type IV collagen) using fluorescently labeled antibodies. Vimentin and α-actin are not detected (b); c — formation of capillary-like structures by ECFC cells on Matrigengel; a, c — phase-contrast microscopy, scale bar — 500 μm; b — confocal microscopy, scale bar — 50 μm.

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4. Fig. 3. Comparison of ECFC colony growth on EGM2 and EndoBoost Plus media: a – photographs of the growth dynamics of the same colonies on different media over 17 days; b – curves of the increase in the number of cells in the ECFC culture over 53 days (cell counting during passages).

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5. Fig. 4. Real-time cell culture growth for 72 h on EGM2, EndoBoost and EndoBoost Plus (xCELLigence): a — curve of the dependence of the cell index (CI) on time; b — CI; c — slope of the curve on different media after 72 h of cultivation.

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