Differentiation of adipocytes and osteocytes from human adipose and placental mesenchymal stem cells

Abstract

Objectives: Mesenchymal stem cells (MSC) can be isolated from adult tissues such as adipose tissue and other sources. Among these sources, adipose tissue (because of easy access) and placenta (due to its immunomodulatory properties, in addition to other useful properties), have attracted more attention in terms of research. The isolation and comparison of MSC from these two sources provides a proper source for clinical experimentation. The aim of this study was to compare the characteristics of MSC isolated from human adipose tissue and placenta.

Materials and methods: Adipose and placental MSC were isolated from the subcutaneous adipose tissues of 10 healthy women (25 to 40 years) and from a fresh term placenta (n= 1), respectively. Stem cells were characterized and compared by flow cytometry using CD29, CD31, CD34, CD44, CD45, CD105, CD166 and HLA-DR markers. Osteocytes and adipocytes were differentiated from isolated human mesenchymal stem cells (HMSC).

Results: Adipose and placenta-derived MSC exhibited the same morphological features. ADSC differentiated faster than placenta; however, both were differentiated, taking up to 21 days for osteocyte and 14 days for adipocyte differentiation. About 90% of PLC-MSC and ADSC were positive for CD29, CD44, CD105, and CD166; and negative for CD31, CD34, CD45, and HLA-DR.

Conclusion: The two sources of stem cells showed similar surface markers, morphology and differentiation potential and because of their multipotency for differentiating to adipocytes and osteocytes, they can be applied as attractive sources of MSC for regenerative medicine.

Keywords: Adult stem cells; Differentiation; Fetal stem cells; Mesenchymal stem cells.

Figure 1

Human mesenchymal stem cells (a-i), Adipogenic and osteogenic differentiation; (g,h) morphology and growth of fibroblastoid-adherent cells or placenta-derived Mesenchymal stem cells at phase 0 on days 2, 5 and 12, respectively (a-c). (a) Small, round cells (Magnification: ×100). (b,c) Long, thin cells (Magnification: ×100) with spindle-shape, which were reached to high confluence and were formed in a colony, as seen in the picture. (c) With morphology and growth of adipose-derived Mesenchymal stem cells at passage 0 on days 2, 4 and 8 (colony), respectively (d-f). (d) A single long spindle-shaped ADSC with thin processes; (Magnification: ×1000). (e,f) ADSCs that were growing (cells were created in a colony (f)) (Magnification: ×100). Adipogenic differentiation was evidenced by the formation of lipid vacuoles (yellow) by Oil Red O staining at passage three in adipose-derived MSC (g) (Magnification: ×1000). Osteogenic differentiation was evidenced by the formation of a mineralized matrix at passage three in adipose-derived MSC (Magnification: ×400) (h) Undifferentiated adipose MSC (i) Scale bar is 50 μm d, g, h) and 100 μm (a, b, c, e, f, i)

Figure 2

Immunophenotyping analysis of adipose mesenchymal stem cells. ADSC was positive for CD105, CD29, CD44 and CD166, and negative for CD31, HLA-DR, CD45 and CD34. Control histogram is highlighted in red. a: Control and CD105; b: Control and CD29; c: Control and CD44; d: Control and CD166; e: Control and CD31; f: Control and HLA DR; g: Control and CD45; h: Control and CD34

Figure 3

Immunophenotyping analysis of placenta mesenchymal stem cells. PLC-MSC was positive for CD105, CD29, CD44 and CD166, and negative for CD31, HL-DR, CD45 and CD34. Control histogram is highlighted in red. a: Control and CD105; b: Control and CD29; c: Control and CD44; d: Control and CD166; e: Control and CD31; f: Control and HLA DR; g: Control and CD45; h: Control and CD34