CD90 Expression on human primary cells and elimination of contaminating fibroblasts from cell cultures

Kisselbach, Lynn; Merges, Michael; Bossie, A; Boyd, Ann L.

doi:10.1007/s10616-009-9190-3

Cited by 116 publications

(94 citation statements)

References 20 publications

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“…When selecting for CAF populations using antibody‐based methods such as FACS, it is essential that multiple surface markers are used in order to avoid any chance of introducing unintentional subtype bias. Other available surface markers such as PDGFRα/β work well here, as do more general fibroblast surface markers like Thy‐1 cell surface antigen (CD90), provided that the cell population is also subjected to selection with negative markers . This is especially important, as a significant number of commonly used fibroblast markers display expression over a number of different cell types, running the risk of inadvertent sample contamination if a proper negative selection is not carried out.…”

Section: Resultsmentioning

confidence: 99%

In search of definitions: Cancer‐associated fibroblasts and their markers

Nurmik

Ullmann

Rodriguez

et al. 2019

Intl Journal of Cancer

470

428

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The tumor microenvironment has been identified as one of the driving factors of tumor progression and invasion. Inside this microenvironment, cancer‐associated fibroblasts (CAFs), a type of perpetually activated fibroblasts, have been implicated to have a strong tumor‐modulating effect and play a key role in areas such as drug resistance. Identification of CAFs has typically been carried based on the expression of various “CAF markers”, such as fibroblast activation protein alpha (FAP) and alpha smooth muscle actin (αSMA), which separates them from the larger pool of fibroblasts present in the body. However, as outlined in this Review, the expression of various commonly used fibroblast markers is extremely heterogeneous and varies strongly between different CAF subpopulations. As such, novel selection methods based on cellular function, as well as further characterizing research, are vital for the standardization of CAF identification in order to improve the cross‐applicability of different research studies in the field. The aim of this review is to give a thorough overview of the commonly used fibroblast markers in the field and their various strengths and, more importantly, their weaknesses, as well as to highlight potential future avenues for CAF identification and targeting.

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Section: Resultsmentioning

confidence: 99%

In search of definitions: Cancer‐associated fibroblasts and their markers

Nurmik

Ullmann

Rodriguez

et al. 2019

Intl Journal of Cancer

470

428

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“…Although these nonmyocyte populations have not been fully characterized, the markers used suggest that they represent a combination of fibroblasts (CD90) 21 , vascular smooth muscle cells (CD140B) 22 and endothelial cells (CD31). Access to enriched populations of each of these cell types together with cardiomyocytes will allow detailed investigation of interactions between them.…”

Section: Discussionmentioning

confidence: 99%

SIRPA is a specific cell-surface marker for isolating cardiomyocytes derived from human pluripotent stem cells

et al. 2011

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To identify cell-surface markers specific to human cardiomyocytes, we screened cardiovascular cell populations derived from human embryonic stem cells (hESCs) against a panel of 370 known CD antibodies. This screen identified the signal-regulatory protein alpha (SIRPA) as a marker expressed specifically on cardiomyocytes derived from hESCs and human induced pluripotent stem cells (hiPSCs), and PECAM, THY1, PDGFRB and ITGA1 as markers of the nonmyocyte population. Cell sorting with an antibody against SIRPA allowed for the enrichment of cardiac precursors and cardiomyocytes from hESC/hiPSC differentiation cultures, yielding populations of up to 98% cardiac troponin T-positive cells. When plated in culture, SIRPA-positive cells were contracting and could be maintained over extended periods of time. These findings provide a simple method for isolating populations of cardiomyocytes from human pluripotent stem cell cultures, and thereby establish a readily adaptable technology for generating large numbers of enriched cardiomyocytes for therapeutic applications.Generation of cardiovascular cells from human pluripotent stem cells (hPSCs) in culture could provide a powerful model system for investigating cellular interactions and molecular regulators that govern the specification, commitment and maturation of these lineages, as well as a unique and unlimited source of human cardiomyocytes for drug testing and regenerative medicine strategies [1][2][3][4] . Translating this potential into practice, however, will depend on the development of technologies that enable the reproducible generation of highly enriched populations of cardiomyocytes, as contaminating cell types could affect drug Reprints and permissions information is available online at

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“…To analyse ORFV‐induced effects on the viability of human skin cells, primary human foreskin‐derived KC or FB were infected with the patient‐derived, replication‐competent ORFV isolate B029 or inactivated ORFV (iORFV) and monitored for morphological cellular changes. The purity of the cell preparation was confirmed by flow cytometric analysis of CD45 and CD90 expression (Figure ). Replication‐competent ORFV rapidly induced morphological changes in KC (Figure A).…”

Section: Resultsmentioning

confidence: 97%

Orf virus infection of human keratinocytes and dermal fibroblasts: Limited virus detection and interference with intercellular adhesion molecule‐1 up‐regulation

Schneider

Protschka

Müller

et al. 2019

Experimental Dermatology

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Orf virus (Parapoxvirus ovis, ORFV) is a dermatotropic virus causing pustular dermatitis in small ruminants and humans. We analysed isolated human primary keratinocytes (KC) and dermal fibroblasts (FB) for cell death and virus replication by infection with a patient‐derived ORFV isolate. ORFV infection was associated with rapid induction of cell death in KC allowing for considerable virus removal. Upon infection with ORFV, KC and FB harboured intracytoplasmic ORFV and showed viral protein presence; however, missing virus spread indicated an abortive infection. Upon ORFV exposure, KC but not FB secreted the pro‐inflammatory cytokine interleukin (IL)‐6. ORFV infection enhanced the frequency of KC expressing intercellular adhesion molecule (ICAM)‐1 which was independent of IL‐6. Interestingly, ORFV inhibited ICAM‐1 up‐regulation on infected but not on non‐infected KC. Even interferon‐γ, a potent inducer of ICAM‐1, up‐regulated ICAM‐1 only on non‐infected KC. Transfer of ORFV‐free supernatant from infected to non‐infected KC induced ICAM‐1 on non‐infected KC pointing to the involvement of soluble mediator(s). Similarly as in KC, in FB interference with ICAM‐1 up‐regulation by ORFV infection was also observed. In conclusion, we shed light on epidermal and dermal defense mechanisms to ORFV infection and point to a novel ICAM‐1‐related immune evasion mechanism of ORFV in human skin.

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CD90 Expression on human primary cells and elimination of contaminating fibroblasts from cell cultures

Cited by 116 publications

References 20 publications

In search of definitions: Cancer‐associated fibroblasts and their markers

In search of definitions: Cancer‐associated fibroblasts and their markers

SIRPA is a specific cell-surface marker for isolating cardiomyocytes derived from human pluripotent stem cells

Orf virus infection of human keratinocytes and dermal fibroblasts: Limited virus detection and interference with intercellular adhesion molecule‐1 up‐regulation

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