Human Mesenchymal Stem Cell Proliferation and Osteogenic Differentiation in Fibrin Gels in Vitro

Catelas, Isabelle; Sese, Nadjah; Wu, Benjamin M.; Dunn, James C.Y.; Helgerson, Sam; Tawil, Bill

doi:10.1089/ten.2006.12.ft-186

Cited by 50 publications

(72 citation statements)

References 34 publications

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“…In that study, the authors used 18S rRNA as normalizer gene based on preliminary assays comparing its stability against GAPDH. Later reports, as the one by Catelas et al (2006), also used 18S rRNA as a normalizer gene under similar experimental conditions. Other studies, as recent as the works by Cho et al (2005), Abdallah et al (2005) and Friedman et al (2006), used ACTB for normalizing the gene expression levels of hBMSCs under osteogenic conditions.…”

Section: Discussionmentioning

confidence: 97%

Housekeeping gene stability influences the quantification of osteogenic markers during stem cell differentiation to the osteogenic lineage

et al. 2010

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Real-time reverse transcription PCR (RT-qPCR) relies on a housekeeping or normalizer gene whose expression remains constant throughout the experiment. RT-qPCR is commonly used for characterization of human bone marrow mesenchymal stem cells (hBMSCs). However, to the best of our knowledge, there are no studies validating the expression stability of the genes used as normalizers during hBMSCs differentiation. This work aimed to study the stability of the housekeeping genes b-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ribosomal protein L13A (RPL13A) during the osteogenic differentiation of hBMSCs. Their stability was evaluated via RT-qPCR in 14 and 20 day differentiation assays to the osteogenic lineage. Different normalization strategies were evaluated to quantify the osteogenic markers collagen type I, bone sialoprotein and osteonectin. Cell differentiation was confirmed via alizarin red staining. The results demonstrated up-regulation of b-actin with maximum fold changes (MFC) of 4.38. GAPDH and RPL13A were not regulated by osteogenic media after 14 days and presented average fold changes lower than 2 in 20 day cultures. RPL13A (MFC \ 2) had a greater stability when normalizing as a function of culture time compared with GAPDH (MFC B 2.2), which resulted in expression patterns of the osteogenic markers more consistent with the observed differentiation process. The results suggest that b-actin regulation could be associated with the morphological changes characteristic of hBMSCs osteogenic differentiation, and provide evidence for the superior performance of RPL13A as a normalizer gene in osteogenic differentiation studies of hBMSCs. This work highlights the importance of validating the normalizer genes used for stem cells characterization via RT-qPCR.

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Section: Discussionmentioning

confidence: 97%

Housekeeping gene stability influences the quantification of osteogenic markers during stem cell differentiation to the osteogenic lineage

et al. 2010

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“…Mineralised tissue was seen to accumulate in the pores that formed in the fibrin gel, which were surrounded by numerous cells (Hou et al 2008). Catelas et al (2006) also showed that MSCs encapsulated in fibrin gels proliferate, show osteogenic differentiation, and secrete mineralised tissue. They, however, observed that the MSCs did not fully differentiate to mature osteoblasts within the 28 days of in vitro culture.…”

Section: Bonementioning

confidence: 93%

Cell encapsulation using biopolymer gels for regenerative medicine

2010

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To cite this version:Nicola C. Hunt, Liam M. Grover. Cell encapsulation using biopolymer gels for regenerative medicine. Biotechnology Letters, Springer Verlag, 2010, 32 (6), pp.733-742. <10.1007/s10529-010-0221-0>. Abstract There has been a consistent increase in the mean life expectancy of the population of the developed world over the past century. Healthy life expectancy, however, has not increased concurrently. As a result we are living a larger proportion of our lives in poor health and there is a growing demand for the replacement of diseased and damaged tissues. While traditionally tissue grafts have functioned well for this purpose, the demand for tissue grafts now exceeds the supply. For this reason, research in regenerative medicine is rapidly expanding to cope with this new demand. There is now a trend towards supplying cells with a material in order to expedite the tissue healing process. Hydrogel encapsulation provides cells with a three dimensional environment similar to that experienced in vivo and therefore may allow the maintenance of normal cellular function in order to produce tissues similar to those found in the body. In this review we discuss biopolymeric gels that have been used for the encapsulation of mammalian cells for tissue engineering applications as well as a brief overview of cell encapsulation for therapeutic protein production. This review focuses on agarose, alginate, collagen, fibrin, hyaluronic acid and gelatin since they are widely used for cell encapsulation. The literature on the regeneration of cartilage, bone, ligament, tendon, skin, blood vessels and neural tissues using these materials has been summarised.

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“…As a whole, all of the effects produced by MSCs here help to solve the problem of chronic hyperinflammation in the wound bed and advance wounds such as diabetic ulcers into the next stages of wound healing. Allogeneic application of MSCs in gelbased products for wound healing holds promise for combatting these issues, as has been done in various applications for MSC therapy using fibrin-based gel systems 68,69 and related mimetics.…”

Section: Msc Immunomodulation In Wound Bedsmentioning

confidence: 99%