Secretion of niche signal molecules in conditions of osteogenic differentiation of multipotent mesenchymal stromal cells induced by textured calcium phosphate coating

: S ispol'zovaniem mul'tipleks-nabora in vitro v kul'ture mul'tipotentnykh mezenkhimnykh stromal'nykh kletok zhirovoĭ tkani cheloveka (MMSK-ZhT) v usloviiakh ee osteogennoĭ differentsirovki, obuslovlennoĭ 14-dnevnym kontaktom s kal'tsiĭ-fosfatnoĭ (KF) poverkhnost'iu razlichnoĭ sherokhovatosti, izuchena sekretsiia riada tsitokinov, khemokinov i faktorov rosta. Dvustoronnee rentgenoamorfnoe KF-pokrytie formirovali na obraztsakh iz kommercheski chistogo titana metodom mikrodugovogo oksidirovaniia v anodnom rezhime. Elektrolit sostoial iz vodnogo rastvora ortofosfornoĭ kisloty (20 mass.%), karbonata kal'tsiia (9 mass.%) i nanoporoshka (diametr chastits 10-30 nm s otdel'nymi agregatami do 100 nm) sinteticheskogo gidroksiapatita (6 mass.%). Posle 4-kh passazheĭ MMSK-ZhT, vydelennye iz lipoaspirata, sokul'tivirovali v konechnoĭ kontsentratsii 1,5´105 zhiznesposobnykh kariotsitov v 1,5 ml standartnoĭ pitatel'noĭ sredy (bez osteogennykh stimuliatorov) s obraztsami v techenie 14 (opredelenie [CD45,34,14,20], CD73, CD90 i CD105 immunofenotipa kletok; analiz sekretornoĭ aktivnosti) i 21 dneĭ (okraska kul'tury alizarinovym krasnym) pri smene kul'tural'noĭ sredy kazhdye 3-4 dnia. Ustanovleno, chto v usloviiakh kontakta in vitro s KF-pokrytiem MMSK-ZhT differentsiruiutsia v osteoblasty, sinteziruiushchie mineralizovannyĭ kostnyĭ matriks. Eto soprovozhdaetsia 2-3-kratnym uvelicheniem doli [CD45,34,14,20]+ krovetvornykh kletok. Signal'nymi molekulami dlia narashchivaniia in vitro gemopoeticheskogo platsdarma v 14-dnevnoĭ differentsiruiushcheĭsia trekhmernoĭ kul'ture MMSK-ZhT iavliaiutsia gumoral'nye faktory krovetvornykh nish: tsitokiny leukemia inhibitory factor (LIF) i stem cell factor (SCF) pri srednem indekse sherokhovatosti KF pokrytiia Ra=2,4-2,6 mkm libo stromal derived factor-1 (SDF-1a (khemokin CXCL12)) pri Ra=3,1-4,4 mkm.