Pilot in vitro study of the parameters of artificial niche for osteogenic differentiation of human stromal stem cell pool

Abstract

The aim of this research is experimental investigation of the topography and evaluation of some parameters of artifi cial microterritories promoting osteogenic differentiation of stromal stem cells. A technique of short-term culturing of prenatal human lung stromal cells with fi broblastoid morphology on calcium phosphate substrates with known topography was used. Judging from secretory activity of the cell culture (osteocalcin, alkaline phosphatase), stromal stem cells directly interacting with calcium phosphate discs have advantage in manifestation of osteoblast-like functional activity in comparison with cells cultured on plastic. Rough surfaces of calcium phosphate discs stimulate the formation of spatial human fi broblastoid cell culture. The cells with positive reaction to acid phosphatase are located on spheroliths forming the relief of calcium phosphate coatings. The cells with positive reaction to alkaline phosphatase (marker of osteoblasts) populate hollows (niches) of the artifi cial surface. The niche for induction of osteogenic differentiation of human multipotent mesenchymal stem cells is apparently a structural and functional formation. It can be characterized by an index calculated as the ratio of the total area occupied by alkaline phosphatase-positive cells to the area of artifi cial surface occupied by one stained cell.

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

“…On the other hand, surface and bulk features ( R a , mass and thickness) are very processible and connected closely with the applied voltages and biological reactions [ 6 , 20 , 37 , 38 , 39 ]. Because some variations of calcium, phosphorus, oxygen, and titanium contents and phase compounds accompanied with surface and bulk indices [ 37 ], microarc CaP-coated samples with a fixed range of R a = 2–5 µm were used in the different series of experiment ( Table 1 , Table 2 , Table 3 and Table 4 , Tables S1–S6 ).…”

Biocompatible nanostructured coatings based on calcium phosphates prepared by means of rf-magnetron sputtering deposition

“…The tested samples were held in 24-well plates (Costar) with the final cells concentration of 3×10 4 viable karyocytes in 1 ml of osteogenous medium. The content of medium, methods used to estimate the morphology and ALP staining of cultivated cells, alkaline phosphatases (ALP) activity in supernatants, optical and scanning electron microscopy (SEM) of samples were applied according to the protocols described in the study [9].…”

Modeling of the mesenchymal stem cell microenvironment as a prospective approach to tissue bioengineering and regenerative medicine (a short review)

“…В ýтом случае микротерритории МСК, как полагают, располагаются вдоль участков ýндоста, в которых есть гипоксическая микросреда. Действительно, наш недавний ýксперимент показал остеогенную дифференциацию и созревание пренатальных стромальных клеток из легких человека (региональный пул МСК) при краткосрочном in vitro контакте с шероховатым скаффолдом КФ, имитирующим минеральное вещество кости [28].…”