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Differentiation of human pluripotent stem cell-derived mesenchymal progenitors into osteogenic, chondrogenic and adipogenic lineages

Alexandra Bergman
Göteborg : Chalmers tekniska högskola, 2012. 54 s.
[Examensarbete på avancerad nivå]

Abstract Differentiated cell types are of great interest in applications such as tissue engineering and drug screening. As harvesting of fully differentiated, adult cells neither is a safe nor scalable alternative, in vitro differentiated cells have become an attractive option. hES-MP002.5 (mesenchymal progenitors) is a cell type established by and at Cellectis Stem Cells from human embryonic stem cells (hESCs), that carries adult mesenchymal stem cell like characteristics. An examination of hES-MP002.5’s mesenchymal differentiation potential was performed in this project, in particular into the osteogenic, chondrogenic and adipogenic lineage. A second objective was to establish a mesenchymal progenitor cell line from a human induced pluripotent stem cell line (ChiPSC4) and compare its characteristics to those of hES-MP002.5. Results were obtained primarily through the use of differentiation in cell culture, histological stains, immunocytochemistry and quantitative PCR. An osteogenic differentiation study concluded that hES-MP002.5 show vast osteogenic potential when a previously established protocol was used. However, hES-MP002.5 showed near to no responsiveness to variations of commonly used chondrogenic or adipogenic treatments in terms of visible chondrogenic extracellular matrix deposition or intracellular lipid accumulation, respectively. Yet, hypoxic culture conditions ought to be considered if chondrogenic differentiation is attempted in future studies. Gene expression studies of adipogenically induced cells implied that a gene essential for adipogenic differentiation was down- regulated for both MP-lines; a fact that could help to explain the low differential response. With an established protocol for hESCs, a cell population resembling hES-MP002.5 was developed from ChiPSC4 (ChiPSC4-MP), but the cell population seemed less homogenous and had lower proliferative potential. hES-MP002.5 and ChiPSC4-MP seemed to respond similarly to mesenchymal differentiation inductions – however ChiPSC4-MP consistently showed a weaker response. This might be due to heterogeneity in the cellular population obtained. With the broad laboratory investigations in mind, it is not recommended that hES-MP002.5 is marketed as a mesenchymal progenitor with the ability of differentiating into chondrogenic and adipogenic lineages. A recommendation is that the protocol for mesenchymal progenitor establishment is reviewed and further developed, since the MP-cells established from both embryonic and induced pluripotent stem cells share the same strengths and weaknesses.

Nyckelord: Stem cell, mesenchymal progenitor, pluripotency, differentiation, osteogenic, chondrogenic, adipogenic, extra cellular matrix

Publikationen registrerades 2013-10-25. Den ändrades senast 2016-12-20

CPL ID: 185646

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