Osteoinduction in Umbilical Cord- and Palate Periosteum-Derived Mesenchymal Stem Cells

Abstract

Adult abdominoplasty (AA) fat is an ideal source for mesenchymal stem cells (MSCs) because it is discarded after surgery, abundant, and easy to harvest. Children however, do not have the same abundant quantities of fat as adults, nor are they likely to undergo a procedure during which fat is routinely discarded. Hence, finding an alternate source for MSCs in children is a reasonable strategy. Two such sources are the palate periosteum (PP) and the umbilical cord (UC). Advantages for PP as a source of MSCs are accessibility during palate repair, ease of harvest, and minimal risk to the patient. The UC, like AA, is a discarded tissue, with a theoretically unlimited supply, which can be harvested in children with craniofacial bone abnormalities in advance of reconstructive procedures. Our objective in this study is to characterize MSCs from 3 sources (AA, PP, and UC) by surface marker prevalence, and to assess osteoinductive capability. Institutional review board approval was obtained for harvest of AA, PP, and UC. The presence of MSCs was determined using immunostaining and flow cytometry for cell surface markers CD73, CD90, CD105, and SSEA-4. Osteogenesis was induced using osteogenic medium. Osteoinduction was evaluated using Alizarin red staining, and real-time polymerase chain reaction for bone morphogenetic protein-2, alkaline phosphatase, and osteocalcin at 7, 14, and 21 days. MSCs from AA, PP, and UC all stained positive for CD73, CD90, CD105, and SSEA-4. Flow cytometry demonstrated significant differences in expression of CD90 and SSEA-4 but similar values for CD73 and CD105. Following osteoinduction, MSCs from all sources stained positive for calcium deposition. In UC MSCs, reverse transcriptase-polymerase chain reaction demonstrated greater elevation in bone morphogenetic protein-2 and alkaline phosphatase mRNA beginning at day 7 and extending to day 21. Osteocalcin mRNA levels were comparable for all 3 sources of MSCs. For children with craniofacial bone defects, UC-derived MSCs may be ideal for tissue engineered bone: temporally, the UC can be harvested in advance of surgical timing for the need for bone, is readily available, easy to harvest, and leads to osteoinduction that is more robust than either AA or PP.

Patterning network structure to spatially control cellular remodeling and stem cell fate within 3-dimensional hydrogels

“…14 days following incubation in 1:1 adipogenic/osteogenic media, hMSC differentiation was assessed by quantitative protein assays for markers of osteogenesis (alkaline phosphatase, ALP) and adipogenesis (oil red O), gene expression analysis, histological staining for ALP and oil red O, and immunostaining for osteocalcin (OC; osteogenic differentiation) and fatty acid binding protein (FABP; adipogenic differentiation). Our choice of differentiation markers was based on literature demonstrating specificity of each marker to its respective lineage at 14 days following hMSC induction [36,37].…”

Juvenile Swine Surgical Alveolar Cleft Model to Test Novel Autologous Stem Cell Therapies

“…These plates are both nonimmunogenic and biodegradable. We have previously demonstrated that these electrospun scaffolds support MSC attachment, migration, proliferation, osteogenic differentiation, and matrix deposition 30,53 ; work by others has demonstrated adequate structural support for treating bone defects in vivo. 54,55 In this study, bone generated using differentiated UC MSCs on PLGA NMFS was better (though not statistically significant) by CT volumetric analysis and histologically similar to the bone generated using cancellous bone from the iliac crest.…”

“…Color images available online at www.liebertpub.com/tec assayed by flow cytometry, as previously described. 30 Passage 0 cells were seeded at a density of 5 · 10 3 cells/cm 2 , grown in Dulbecco's modified Eagle's medium (DMEM) 15% fetal bovine serum (FBS) and sub-passaged at 80% confluence.…”

Comparison of Different Sources of Mesenchymal Stem Cells: Palatal versus Lipoaspirated Adipose Tissue

“…Stem cells have also been isolated from new, more accessible sources, including gingiva [Yang et al, 2013], oral mucosa, palatal connective tissue [Roman et al, 2013], and palatal periosteum [Caballero et al, 2010]. The main goal of the present study was to characterize and compare a novel stem cell population obtained from PAT-and LAT-derived stem cells.…”