Detection In Vitro and Quantitative Estimation of Artificial Microterritories Which Promote Osteogenic Differentiation and Maturation of Stromal Stem Cells
Extracellular matrix can regulate multipotent mesenchymal stromal cells (MMSC) differentiation, with potential applications for tissue engineering. A relief of mineralized bone takes essential effect on MMSC fate. Nevertheless, delicate structure and a hierarchy of niches for stromal stem cells and its quantitative parameters are not practically known. Here, we describe the protocol to define the basic approach providing a guiding for in vitro identification of quantitative features of artificial calcium phosphate niches which promotes osteogenic differentiation and maturation of stromal stem cell.
Key wordsPrenatal stromal cells of human lung Calcium phosphate surface Short-term culture Artificial osteogenic niches Alkaline phosphatase Acid phosphatase Osteocalcin Immunocytochemistry Light microscopy Scanning electron microscopy
The authors are deeply indebted to: professor Yu.P. Sharkeev and E.V. Legostaeva Ph.D. (Institute of Strength Physics and Materials Science, SB of RAS, Tomsk, Russia) for designing and digital imaging of titanium specimens with calcium phosphate coating; K.V. Zaitsev Ph.D. (Stem Cells Bank Ltd., Tomsk, Russia) for cell culture provision; Joint Use Center for Materials Science (Tomsk State University, Tomsk, Russia) for microscopic equipment use.
This work was supported by the Federal Goal Program of Russian Federation (grant No 8036).
- 14.Khlusov IA, Khlusova M Yu, Zaitsev KV et al (2011) Pilot in vitro study of the parameters of artificial niche for osteogenic differentiation of human stromal stem cell pool. Bull Exp Biol Med 150:535–542Google Scholar
- 15.Burtis CA, Ashwood ER (eds) (2001) Tietz fundamentals of clinical chemistry, 5th edn. W.B. Saunders Company, PhiladelphiaGoogle Scholar
- 16.Sharkeev Yu P, Legostaeva EV, Eroshenko A Yu et al (2009) The structure and physical and mechanical properties of a novel biocomposite material, nanostructured titanium-calcium-phosphate coating. Compos Interfac 16:535–546Google Scholar
- 19.da Silva Meirelles L, Chagastelles PC, Nardi NB (2006) Mesenchymal stem cells reside in virtually all post-natal organs and tissues. JCS 119:2204–2213Google Scholar
- 21.Riggs BL, Melton LJ III (1996) Osteoporosis. Etiology, diagnosis, and management, 2nd edn. Lippincott-Raven, Philadelphia, New YorkGoogle Scholar
- 23.Khlusov IA, Dekhtyar Yu, Khlusova M Yu et al (2013) Novel concepts of “Niche-Relief” and “Niche-Voltage” for stem cells as a base of bone and hematopoietic tissues biomimetic engineering. IFMBE Proc 38:99–102. doi: 10.1007/978-3-642-34197-7_26