scholarly journals Mechanical Characterization of Differentiated Human Embryonic Stem Cells

2009 ◽  
Vol 131 (6) ◽  
Author(s):  
Gidon Ofek ◽  
Vincent P. Willard ◽  
Eugene J. Koay ◽  
Jerry C. Hu ◽  
Patrick Lin ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Tissue Engineering ◽  
Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Articular Chondrocytes ◽  
Embryonic Stem ◽  
Density Separation ◽  
Undifferentiated Hescs ◽  
Cell Source ◽  
Tgf Β1

Human embryonic stem cells (hESCs) possess an immense potential in a variety of regenerative applications. A firm understanding of hESC mechanics, on the single cell level, may provide great insight into the role of biophysical forces in the maintenance of cellular phenotype and elucidate mechanical cues promoting differentiation along various mesenchymal lineages. Moreover, cellular biomechanics can provide an additional tool for characterizing stem cells as they follow certain differentiation lineages, and thus may aid in identifying differentiated hESCs, which are most suitable for tissue engineering. This study examined the viscoelastic properties of single undifferentiated hESCs, chondrogenically differentiated hESC subpopulations, mesenchymal stem cells (MSCs), and articular chondrocytes (ACs). hESC chondrogenesis was induced using either transforming growth factor-β1(TGF-β1) or knock out serum replacer as differentiation agents, and the resulting cell populations were separated based on density. All cell groups were mechanically tested using unconfined creep cytocompression. Analyses of subpopulations from all differentiation regimens resulted in a spectrum of mechanical and morphological properties spanning the range of hESCs to MSCs to ACs. Density separation was further successful in isolating cellular subpopulations with distinct mechanical properties. The instantaneous and relaxed moduli of subpopulations from TGF-β1 differentiation regimen were statistically greater than those of undifferentiated hESCs. In addition, two subpopulations from the TGF-β1 group were identified, which were not statistically different from native articular chondrocytes in their instantaneous and relaxed moduli, as well as their apparent viscosity. Identification of a differentiated hESC subpopulation with similar mechanical properties as native chondrocytes may provide an excellent cell source for tissue engineering applications. These cells will need to withstand any mechanical stimulation regimen employed to augment the mechanical and biochemical characteristics of the neotissue. Density separation was effective at purifying distinct populations of cells. A differentiated hESC subpopulation was identified with both similar mechanical and morphological characteristics as ACs. Future research may utilize this cell source in cartilage regeneration efforts.

Embryonic Stem Cells as a Cell Source for Tissue Engineering

2007 ◽  
pp. 445-458
Author(s):  
Ali Khademhosseini ◽  
Jeffrey M. Karp ◽  
Sharon Gerecht ◽  
Lino Ferreira ◽  
Gordana Vunjak-Novakovic ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Cell Source ◽  
A Cell

scholarly journals Tissue Engineering of Vascularized Cardiac Muscle From Human Embryonic Stem Cells

2007 ◽  
Vol 100 (2) ◽  
pp. 263-272 ◽  
Author(s):  
Oren Caspi ◽  
Ayelet Lesman ◽  
Yaara Basevitch ◽  
Amira Gepstein ◽  
Gil Arbel ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Cardiac Muscle ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem

Derivation of SSEA4-/CD73+ Mesenchymal Stem Cells from Human Embryonic Stem Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2579-2579
Author(s):  
Parul Trivedi ◽  
Peiman Hematti
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Clinical Applications ◽  
Differentiation Potential ◽  
Hla Class I Antigens ◽  
Undifferentiated Hescs

Abstract Human embryonic stem cells (hESCs) could potentially provide a renewable source of different types of cells for cell therapy applications. Recently, mesenchymal stem cells (MSCs) have been derived from hESCs either through co-culturing with murine OP9 bone marrow stromal cell line or directly from hESCs without co-culturing with OP9 cells. Although the latter methodology is clinically advantageous over co-culturing with an animal cell layer those mesenchymal cells were reported to be positive for SSEA4. SSEA4 is a marker of undifferentiated hESCs and thus the presence of this marker on hESC-derived cells could potentially be problematic for clinical applications. We have recently achieved a novel and reproducible methodology for deriving a pure population of SSEA4-/CD73+ MSCs from federally approved hESC lines H1 and H9. To initiate the differentiation of hESCs to MSCs, we cultured undifferentiated hESCs on matrigel plates in murine embryonic fibroblast conditioned media with media changes every 3 days. Under these culture conditions a portion of embryonic stem cells differentiated into fibroblast looking cells. Through a multi-step process which involved the use of a culture methodology similar to what is being used to culture bone marrow (BM)-derived MSCs, and passaging cultured cells at defined time points we were able to derive a pure population of cells that were uniformly positive for MSC marker CD73 in about a 4-weeks period. These cells had fibroblast/mesenchymal looking morphology, and expressed cell surface marker antigens similar to what has been reported for adult human BM-derived MSCs: they are positive for CD29, CD44, CD54, CD71, CD90, glycophorin A, CD105, and were negative for hematopoietic markers such as CD34 and CD45. Similar to adult BM-derived MSCs these cells express HLA class-I antigens but not class-II antigens. Using established differentiation protocols we could differentiate the hESC-derived CD73+ MSCs into adipocytes, osteocytes, and chondrocytes as verified by immunohistochemistry and RT-PCR assays. So far we have grown these CD73+ MSCs up to passages 15–18. These cells retained their differentiation potential, and were karotypically normal when tested at passage 12. Most importantly, we did not observe any MSCs that were double positive for CD73 and SSEA4 antigen at any time point during our experiments. MSCs from a variety of fetal and adult sources are in various stages of clinical trials with some encouraging preliminary results. Our hESC-derived MSCs that are very similar to adult BM-derived MSCs regarding their growth and morphologic properties, immunophenotypic characteristics, differentiation potential, and importantly are devoid of hESC marker SSEA4 could potentially provide a novel source of MSCs for clinical applications.

scholarly journals Tissue Engineering with Chondrogenically Differentiated Human Embryonic Stem Cells

Stem Cells ◽  
2007 ◽  
Vol 25 (9) ◽  
pp. 2183-2190 ◽  
Author(s):  
Eugene J. Koay ◽  
Gwen M. B. Hoben ◽  
Kyriacos A. Athanasiou
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem

scholarly journals Red Ginseng Extract Facilitates the Early Differentiation of Human Embryonic Stem Cells into Mesendoderm Lineage

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Yoon Young Kim ◽  
Seung-Yup Ku ◽  
Zev Rosenwaks ◽  
Hung Ching Liu ◽  
Sun Kyung Oh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Embryoid Bodies ◽  
Red Ginseng ◽  
Cardiac Progenitors ◽  
Early Differentiation ◽  
Undifferentiated Hescs ◽  
Wide Range

Human embryonic stem cells (hESCs) have capacities to self-renew and differentiate into all cell typesin vitro. Red ginseng (RG) is known to have a wide range of pharmacological effectsin vivo; however, the reports on its effects on hESCs are few. In this paper, we tried to demonstrate the effects of RG on the proliferation and differentiation of hESCs. Undifferentiated hESCs, embryoid bodies (EBs), and hESC-derived cardiac progenitors (CPs) were treated with RG extract at 0.125, 0.25, and 0.5 mg/mL. After treatment of undifferentiated hESCs from day 2 to day 6 of culture, BrdU labeling showed that RG treatment increased the proliferation of hESCs, and the expression of Oct4 and Nanog was increased in RG-treated group. To find out the effects of RG on early differentiation stage cells, EBs were treated with RG extract for 10 days and attached for further differentiation. Immunostaining for three germ layer markers showed that RG treatment increased the expressions of Brachyury and HNF3βon EBs. Also, RG treatment increased the expression of Brachyury in early-stage and of Nkx2.5 in late-stage hESC-derived CPs. These results demonstrate facilitating effects of RG extract on the proliferation and early differentiation of hESC.

scholarly journals A scaffold-free approach to cartilage tissue generation using human embryonic stem cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Lauren A. Griffith ◽  
Katherine M. Arnold ◽  
Bram G. Sengers ◽  
Rahul S. Tare ◽  
Franchesca D. Houghton
Keyword(s):  
Stem Cells ◽  
Articular Cartilage ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Articular Chondrocytes ◽  
Embryonic Stem ◽  
Cartilage Tissue ◽  
Human Articular Cartilage ◽  
3D Scaffold ◽  
Tissue Constructs

AbstractArticular cartilage functions as a shock absorber and facilitates the free movement of joints. Currently, there are no therapeutic drugs that promote the healing of damaged articular cartilage. Limitations associated with the two clinically relevant cell populations, human articular chondrocytes and mesenchymal stem cells, necessitate finding an alternative cell source for cartilage repair. Human embryonic stem cells (hESCs) provide a readily accessible population of self-renewing, pluripotent cells with perceived immunoprivileged properties for cartilage generation. We have developed a robust method to generate 3D, scaffold-free, hyaline cartilage tissue constructs from hESCs that are composed of numerous chondrocytes in lacunae, embedded in an extracellular matrix containing Type II collagen, sulphated glycosaminoglycans and Aggrecan. The elastic (Young’s) modulus of the hESC-derived cartilage tissue constructs (0.91 ± 0.08 MPa) was comparable to full-thickness human articular cartilage (0.87 ± 0.09 MPa). Moreover, we have successfully scaled up the size of the scaffold-free, 3D hESC-derived cartilage tissue constructs to between 4.5 mm and 6 mm, thus enhancing their suitability for clinical application.

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