Quantitative identification of teratoma tissues formed by human embryonic stem cells with TeratomEye

Oh, Steve; Chua, Pauline; Foon, Koh Lin; Ng, E. T. L.; Chin, Angela; Choo, Andre

doi:10.1007/s10529-009-9928-1

Cited by 8 publications

(8 citation statements)

References 6 publications

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“…After 3-4 weeks, the tumors were removed, fixed in 4 % formalin, and embedded in paraffin. The histology of in vivo differentiated three germ layers -respiratory (endoderm), cartilage (mesoderm), and neural rosettes (ectoderm) -was analyzed after hematoxylin and eosin staining [46,47].…”

Section: Pluripotency Analysis After Differentiation In Vitro and In mentioning

confidence: 99%

Pluripotent stem cells as a source of osteoblasts for bone tissue regeneration

et al. 2019

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Appropriate and abundant sources of bone-forming osteoblasts are essential for bone tissue engineering. Pluripotent stem cells can self-renew and thereby offer a potentially unlimited supply of osteoblasts, a significant advantage over other cell sources. We generated mouse embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) from transgenic mice expressing rat 2.3 kb type I collagen promoter-driven green fluorescent protein (Col2.3GFP), a reporter of the osteoblast lineage. We demonstrated that Col2.3GFP ESCs and iPSCs can be successfully differentiated to osteoblast lineage cells that express Col2.3GFP in vitro. We harvested GFP osteoblasts differentiated from ESCs. Genome wide gene expression profiles validated that ESC- and iPSC-derived osteoblasts resemble calvarial osteoblasts, and that Col2.3GFP expression serves as a marker for mature osteoblasts. Our results confirm the cell identity of ESC- and iPSC-derived osteoblasts and highlight the potential of pluripotent stem cells as a source of osteoblasts for regenerative medicine.

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Section: Pluripotency Analysis After Differentiation In Vitro and In mentioning

confidence: 99%

Pluripotent stem cells as a source of osteoblasts for bone tissue regeneration

et al. 2019

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“…Qualitative analyses such as these are more subjective by nature and coupled with the variable and haphazard nature of the EBs/teratomas, these qualitative analyses can only really be used to ascertain trends in differentiation rather than directly comparing the strength of differentiation. While some attempts to more quantitatively assess the histology of teratomas have been made ( Oh et al, 2009 ; Bhagavatula et al, 2010 ), ultimately quantitative gene expression analyses will provide more objective data as to the strength of differentiation. These simple morphogen studies lay the foundations for more nuanced differentiation experiments, that could involve exposing the 3D cultures to a series of alternative morphogens at different times and concentrations to simulate developmental processes.…”

Section: Discussionmentioning

confidence: 99%

Using Advanced Cell Culture Techniques to Differentiate Pluripotent Stem Cells and Recreate Tissue Structures Representative of Teratoma Xenografts

Smith

Aguilar

Owens

et al. 2021

Front. Cell Dev. Biol.

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Various methods are currently used to investigate human tissue differentiation, including human embryo culture and studies utilising pluripotent stem cells (PSCs) such as in vitro embryoid body formation and in vivo teratoma assays. Each method has its own distinct advantages, yet many are limited due to being unable to achieve the complexity and maturity of tissue structures observed in the developed human. The teratoma xenograft assay allows maturation of more complex tissue derivatives, but this method has ethical issues surrounding animal usage and significant protocol variation. In this study, we have combined three-dimensional (3D) in vitro cell technologies including the common technique of embryoid body (EB) formation with a novel porous scaffold membrane, in order to prolong cell viability and extend the differentiation of PSC derived EBs. This approach enables the formation of more complex morphologically identifiable 3D tissue structures representative of all three primary germ layers. Preliminary in vitro work with the human embryonal carcinoma line TERA2.SP12 demonstrated improved EB viability and enhanced tissue structure formation, comparable to teratocarcinoma xenografts derived in vivo from the same cell line. This is thought to be due to reduced diffusion distances as the shape of the spherical EB transforms and flattens, allowing for improved nutritional/oxygen support to the developing structures over extended periods. Further work with EBs derived from murine embryonic stem cells demonstrated that the formation of a wide range of complex, recognisable tissue structures could be achieved within 2–3 weeks of culture. Rudimentary tissue structures from all three germ layers were present, including epidermal, cartilage and epithelial tissues, again, strongly resembling tissue structure of teratoma xenografts of the same cell line. Proof of concept work with EBs derived from the human embryonic stem cell line H9 also showed the ability to form complex tissue structures within this system. This novel yet simple model offers a controllable, reproducible method to achieve complex tissue formation in vitro. It has the potential to be used to study human developmental processes, as well as offering an animal free alternative method to the teratoma assay to assess the developmental potential of novel stem cell lines.

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“…To these ends, the ability to automatically identify and segment specific tissue types and reconstruct them in three dimensions by computerized means would greatly advance this area of developmental biology. Recently, our group and others have made advances in the application of algorithms and techniques used in automated image analysis specifically applied to recognition of tissue types in teratomas[26, 27]. The use of numerous immunohistochemical markers would aid quantitative analysis of specific tissue types (and hence germ layer derivation), although we suspect that some tissues, particularly more undifferentiated and primitive tissues may not have commercially available antibodies that would both perform adequately in formalin-fixed paraffin embedded tissues and also elucidate specific phenotypes.…”

Section: Discussionmentioning

confidence: 99%

Semiquantitative histopathology and 3D magnetic resonance microscopy as collaborative platforms for tissue identification and comparison within teratomas derived from pedigreed primate embryonic stem cells

et al. 2010

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Background Teratoma formation in xenografts is a sufficiently stringent pluripotency assay for stem cells. However, little is known about composition and spatial relationships of tissues within teratomas that may provide clues about development and platforms for studying organ development. Additionally, teratoma formation and analysis lack standards for reporting as assays of pluripotency. Methods Three of 27 total teratomas derived from pedigreed primate embryonic stem cells underwent quantitative 3-dimensional high-resolution magnetic resonance microscopy (MRM). Teratomas were subsequently serially sectioned and tissue types identified, semiquantitated, and correlated with MRM images. Results All teratomas demonstrated tissue derivatives from the three germ layers and approximately 23 different tissue types were identified. Certain tissue groups attempted to form organs more frequently (e.g. trachea/bronchi, small intestine). MRM discriminated some tissues readily (e.g. bone, adipose, cartilage) while other tissue types with like MR intensities could not be distinguished. Conclusion Semiquantitative histopathological analysis of teratomas demonstrates the ability to delineate multiple tissues as derived from ectoderm, mesoderm, or endoderm and to use this information for comparison to other teratomas. MRM provides rapid quantitative imaging of intact teratomas that complements histology and identifies sites of interest for additional biological studies.

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Quantitative identification of teratoma tissues formed by human embryonic stem cells with TeratomEye

Cited by 8 publications

References 6 publications

Pluripotent stem cells as a source of osteoblasts for bone tissue regeneration

Pluripotent stem cells as a source of osteoblasts for bone tissue regeneration

Using Advanced Cell Culture Techniques to Differentiate Pluripotent Stem Cells and Recreate Tissue Structures Representative of Teratoma Xenografts

Semiquantitative histopathology and 3D magnetic resonance microscopy as collaborative platforms for tissue identification and comparison within teratomas derived from pedigreed primate embryonic stem cells

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