Impact of the source and serial passaging of goat mesenchymal stem cells on osteogenic differentiation potential: implications for bone tissue engineering

Abstract

BackgroundAdult mesenchymal stem cells (MSCs) can be conveniently sampled from bone marrow, peripheral blood, muscle, adipose and connective tissue, harvested from various species, including, rodents, dogs, cats, horses, sheep, goats and human beings. The MSCs isolated from adult tissues vary in their morphological and functional properties. These variations are further complicated when cells are expanded by passaging in culture. These differences and changes in MSCs must be considered prior to their application in the clinic or in a basic research study. Goats are commonly used as animal models for bone tissue engineering to test the potential of stem cells for bone regeneration. As a result, goat MSCs isolated from bone marrow or adipose tissue should be evaluated using in vitro assays, prior to their application in a tissue engineering project.ResultsIn this study, we compared the stem cell properties of MSCs isolated from goat bone marrow and adipose tissue. We used quantitative and qualitative assays with a focus on osteogenesis, including, colony forming unit, rate of cell proliferation, tri-lineage differentiation and expression profiling of key signal transduction proteins to compare MSCs from low and high passages. Primary cultures generated from each source displayed the stem cell characteristics, with variations in their osteogenic potentials. Most importantly, low passaged bone marrow MSCs displayed a significantly higher and superior osteogenic potential, and hence, will be the preferred choice for bone tissue engineering in future in vivo experiments. In the bone marrow MSCs, this process is potentially mediated by the p38 MAPK pathway. On the other hand, osteogenic differentiation in the adipose tissue MSCs may involve the p44/42 MAPK pathway.ConclusionsBased on these data, we can conclude that bone marrow and fat-derived MSCs undergo osteogenesis via two distinct signaling pathways. Even though the bone marrow MSCs are the preferred source for bone tissue engineering, the adipose tissue MSCs are an attractive alternative source and undergo osteo-differentiation differently from the bone marrow MSCs and hence, might require a cell-based enhancer/inducer to improve their osteogenic regenerative capacity.

<p>Functionalized Graphene Nanoparticles Induce Human Mesenchymal Stem Cells to Express Distinct Extracellular Matrix Proteins Mediating Osteogenesis</p>

“…The accumulation of calcium in hMSCs was quantitated by the elution of alizarin red dye with 10% cetylpyridinium chloride and the color was read at 570 nm. 42 Background readings due to the substrates alone without any cells were subtracted from the sample readings to eliminate nonspecific values. Data were plotted and statistics performed in Prism (Graphpad).…”

Pencirian dan Pembezaan Osteogenik Sel Stem Pulpa Gigi Kekal dan Gigi Desiduos Manusia

“…Sel stem sumsum tulang (BMSC) dan sel stem adipos (ASC) merupakan sumber MSC yang paling mudah diperoleh kerana mempunyai jumlah sel stem yang banyak. Namun begitu, BMSC menunjukkan tahap komitmen yang lebih tinggi untuk membeza kepada asalan kondrogenik dan osteogenik yang menjadikan BMSC lebih baik berbanding ACS (Elkhenany et al 2016;Xu et al 2017). Walaupun BMSC telah dikaji dengan lebih terperinci, namun disebabkan oleh faktor kesan morbiditi kepada penderma, kesukaran memperoleh sel yang mencukupi dan kehilangan fenotip sel semasa pengkulturan menyebabkan sumber alternatif bagi sel stem daripada sumber organ dan tisu yang lain diperlukan seperti darah periferi (Muhammad Dain et al 2011;Ruzanna et al 2012;.…”

Cellularizing hydrogel-based scaffolds to repair bone tissue: How to create a physiologically relevant micro-environment?

“… 182 Such modifications seem to affect the capacity of MSCs, once seeded into a 3D matrix and implanted in host tissues, to differentiate into osteoblasts and, most importantly, to efficiently produce a bone matrix. 183 Interestingly, treatment of cells with melatonin during this expansion period has been proposed to preserve their differentiation capacity. 184 Given the loss of performances of MSCs upon long-term culture, it is advisable to reduce the number of passages (usually MSCs undergo less than five passages before engraftment), although a compromise must be found between a large number of cells, requiring prolonged amplification and maintenance of therapeutic efficiency of cells.…”