Cell-Secreted Matrices Enhance Osteogenesis in Adipose-Derived Stem Cells Undergoing Lineage Specification

Author(s):  
Heather B. Coan ◽  
Thaleia Teli ◽  
Christoper Booth ◽  
Mark O. Lively ◽  
Mark Van Dyke

Osteogenic differentiation of mesenchymal stem cells (MSC) is important in the field of bone tissue engineering. The identification of biological factors that influence osteogenesis is vital for developing a broader understanding of how complex microenvironments play a role in differentiation. The aim of this study was to demonstrate that adipose-derived stem cell (ADSC) osteogenesis is enhanced through interaction with extracellular matrices (ECM) secreted by ADSC undergoing osteogenesis. ADSC were obtained from human patients following elective abdominoplasty. Cells were selected for plastic adherence, characterized, and induced to differentiate using osteogenic supplements (OS; dexamethasone, ascorbic acid, and beta-glycerol phosphate). Cells were removed at several time points during osteogenesis and the secreted ECM was isolated. Undifferentiated cells were re-seeded onto the cell secreted ECMs and induced to differentiate with OS. At several time points, cells cultured on ECMs or tissue culture plastic controls (i.e. uncoated surface) were collected and RNA isolated. QPCR and gene array analysis revealed enrichment of osteogenic markers and more rapid progression through osteogenic maturational phases in cells seeded onto ECM secreted at the midpoint in differentiation (ca. 15 days). Our results demonstrate that the cumulative deposition of ECM reaches a critical point at approximately 15 days, before which there appear to be no definitive osteogenic cues from the matrix, and after which, strong drivers of osteogenesis are present. The creation of microenvironments that contain essential morphogenic matrix signals is an important step towards methods of growing and differentiating MSC in a rapid effective manner, particularly for bone-related clinical applications.

2018 ◽  
Vol 34 (2) ◽  
pp. 209-223
Author(s):  
Shideh Shaneh ◽  
Fatemeh Shokrolahi ◽  
Parvin Shokrollahi ◽  
Hamid Yeganeh ◽  
Hossein Omidian

Poly(urethane urea) scaffolds were fabricated through combined salt leaching and solvent casting methods. The scaffolds were then functionalized via aminolysis with poly(ethylene glycol) (PEG- g-PUU). To compare its bioactivity, gelatin was also grafted onto the aminolyzed poly(urethane urea) surface (Gel- g-PUU). Chemical changes at the surface were then monitored using quantitative/qualitative methods. Grafting with both gelatin and poly(ethylene glycol) remarkably enhanced the wettability of poly(urethane urea). Proliferation of human adipose–derived mesenchymal stem cells on poly(urethane urea) and the modified poly(urethane urea)s was evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay. The cell experiment results showed that both the modified poly(urethane urea)s enhanced the attachment and proliferation of human adipose–derived mesenchymal stem cells compared to pure poly(urethane urea). Based on previous reports, while a supportive role is observed at adequate poly(ethylene glycol) graft densities, cell adhesion and proliferation are inhibited at very high grafting densities. To correlate the cell data to poly(ethylene glycol) conformations, the surface tension was measured. Data on human adipose–derived mesenchymal stem cells’ attachment/proliferation and contact angle/surface free energy together showed that the grafting density of poly(ethylene glycol) was regulated by optimizing aminolysis conditions, careful selection of poly(ethylene glycol)’s molecular weight, and bulk properties of the matrix poly(urethane urea). As a result, surface overcrowding and brush conformation of the poly(ethylene glycol) chains were avoided, and human adipose–derived mesenchymal stem cell attachment and proliferation occurred on the PEG- g-PUU scaffold at a comparable level to the Gel- g-PUU.


2009 ◽  
Vol 3 (1) ◽  
pp. 61-65 ◽  
Author(s):  
Ana Armiñán ◽  
Carolina Gandía ◽  
José Manuel García-Verdugo ◽  
Elisa Lledó ◽  
José Luis Mullor ◽  
...  

2011 ◽  
Vol 8 (2) ◽  
pp. 200-213 ◽  
Author(s):  
Kian Peng Koh ◽  
Akiko Yabuuchi ◽  
Sridhar Rao ◽  
Yun Huang ◽  
Kerrianne Cunniff ◽  
...  

2017 ◽  
Vol 4 (4) ◽  
pp. 533-542 ◽  
Author(s):  
Guangdun Peng ◽  
Patrick P. L. Tam ◽  
Naihe Jing

Abstract Establishment of progenitor cell populations and lineage diversity during embryogenesis and the differentiation of pluripotent stem cells is a fascinating and intricate biological process. Conceptually, an understanding of this developmental process provides a framework to integrate stem-cell pluripotency, cell competence and differentiating potential with the activity of extrinsic and intrinsic molecular determinants. The recent advent of enabling technologies of high-resolution transcriptome analysis at the cellular, population and spatial levels proffers the capability of gaining deeper insights into the attributes of the gene regulatory network and molecular signaling in lineage specification and differentiation. In this review, we provide a snapshot of the emerging enabling genomic technologies that contribute to the study of development and stem-cell biology.


Author(s):  
Jiri Kalabis ◽  
Kenji Oyama ◽  
Takaomi Okawa ◽  
Hiroshi Nakagawa ◽  
Carmen Z. Michaylira ◽  
...  

2020 ◽  
Author(s):  
Nathalia Azevedo ◽  
Elisa Bertesago ◽  
Ismail Ismailoglu ◽  
Michael Kyba ◽  
Michihiro Kobayashi ◽  
...  

AbstractThe in vitro generation from pluripotent stem cells (PSCs) of different blood cell types, in particular those that are not replenished by hematopoietic stem cells (HSCs) like fetal-derived tissue-resident macrophages and innate-like lymphocytes, is of a particular interest. In order to succeed in this endeavor, a thorough understanding of the pathway interplay promoting lineage specification for the different blood cell types is needed. Notch signaling is essential for the HSC generation and their derivatives, but its requirement for tissue-resident immune cells is unknown. Using mouse embryonic stem cells (mESCs) to recapitulate murine embryonic development, we have studied the requirement for Notch signaling during the earliest B-lymphopoiesis and found that Rbpj-deficient mESCs are able to generate B-1 cells. Their Notch-independence was confirmed in ex vivo experiments using Rbpj-deficient embryos. In addition, we found that upregulation of Notch signaling was needed for the emergence of B-2 lymphoid cells. Taken together, these findings indicate that control of Notch signaling dosage is critical for the different B-cell lineage specification and provides pivotal information for their in vitro generation from PSCs for therapeutic applications.


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