Sterile and disposable fluidic subsystem suitable for clinical high speed fluorescence‐activated cell sorting

Jayasinghe, Sachintha; Wunderlich, Joshua P.; McKee, Angel; Newkirk, Heather L.; Pope, Stephen B.; Zhang, Jiwang; Li, Linheng; Haug, Jeffrey S.

doi:10.1002/cyto.b.20111

Cited by 12 publications

(8 citation statements)

References 37 publications

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“…In addition, preparative sorting of any product infused into patients has to be performed using Good Manufacturing Practices (GMP). GMPs help assure product quality largely by preventing contamination through specific activities such as environmental conditions (e.g., clean room), decontamination practices and testing (23, 62, 63).…”

Section: Conclusion and Novel Applicationsmentioning

confidence: 99%

International Society for Analytical Cytology Biosafety Standard for Sorting of Unfixed Cells

et al. 2007

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Background: Cell sorting of viable biological specimens has become very prevalent in laboratories involved in basic and clinical research. As these samples can contain infectious agents, precautions to protect instrument operators and the environment from hazards arising from the use of sorters are paramount. To this end the International Society of Analytical Cytology (ISAC) took a lead in establishing biosafety guidelines for sorting of unfixed cells (Schmid et al., Cytometry 1997;28:99–117). During the time period these recommendations have been available, they have become recognized worldwide as the standard practices and safety precautions for laboratories performing viable cell sorting experiments. However, the field of cytometry has progressed since 1997, and the document requires an update. Methods: Initially, suggestions about the document format and content were discussed among members of the ISAC Biosafety Committee and were incorporated into a draft version that was sent to all committee members for review. Comments were collected, carefully considered, and incorporated as appropriate into a draft document that was posted on the ISAC web site to invite comments from the flow cytometry community at large. The revised document was then submitted to ISAC Council for review. Simultaneously, further comments were sought from newly‐appointed ISAC Biosafety committee members. Results: This safety standard for performing viable cell sorting experiments was recently generated. The document contains background information on the biohazard potential of sorting and the hazard classification of infectious agents as well as recommendations on (1) sample handling, (2) operator training and personal protection, (3) laboratory design, (4) cell sorter set‐up, maintenance, and decontamination, and (5) testing the instrument for the efficiency of aerosol containment. Conclusions: This standard constitutes an updated and expanded revision of the 1997 biosafety guideline document. It is intended to provide laboratories involved in cell sorting with safety practices that take into account the enhanced hazard potential of high‐speed sorting. Most importantly, it states that droplet‐based sorting of infectious or hazardous biological material requires a higher level of containment than the one recommended for the risk group classification of the pathogen. The document also provides information on safety features of novel instrumentation, new options for personal protective equipment, and recently developed methods for testing the efficiency of aerosol containment. Published 2007 Wiley‐Liss, Inc.

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Section: Conclusion and Novel Applicationsmentioning

confidence: 99%

International Society for Analytical Cytology Biosafety Standard for Sorting of Unfixed Cells

et al. 2007

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“…Consequently, precise isolation of certain cell types, such as shoot apical meristem cells, which are deeply embedded within the embryo, is a considerable challenge. Evidently, FACS in combination with gene expression analysis has been broadly employed for many studies, such as purification of Drosophila melanogaster embryonic cell populations (Cumberledge and Krasnow, 1994;Shigenobu et al, 2006), clinical applications (Jayasinghe et al, 2006;Jaye et al, 2012) and isolation of different cell types in Arabidopsis root and shoot tissue (Birnbaum et al, 2003;De Smet et al, 2008;Yadav et al, 2014). Most of the FACS studies in plants were based on the generation of protoplasts from easily accessible tissues and therefore this method is very difficult to apply to Arabidopsis embryos, in particular in large amounts.…”

Section: Introductionmentioning

confidence: 99%

Cell type-specific transcriptome analysis in the early Arabidopsis thaliana embryo

et al. 2014

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In multicellular organisms, cellular differences in gene activity are a prerequisite for differentiation and establishment of cell types. In order to study transcriptome profiles, specific cell types have to be isolated from a given tissue or even the whole organism. However, wholetranscriptome analysis of early embryos in flowering plants has been hampered by their size and inaccessibility. Here, we describe the purification of nuclear RNA from early stage Arabidopsis thaliana embryos using fluorescence-activated nuclear sorting (FANS) to generate expression profiles of early stages of the whole embryo, the proembryo and the suspensor. We validated our datasets of differentially expressed candidate genes by promoter-reporter gene fusions and in situ hybridization. Our study revealed that different classes of genes with respect to biological processes and molecular functions are preferentially expressed either in the proembryo or in the suspensor. This method can be used especially for tissues with a limited cell population and inaccessible tissue types. Furthermore, we provide a valuable resource for research on Arabidopsis early embryogenesis.

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“…The identification and selection of stem cells within a given tissue/organ largely relies on the presence of specific cell surface markers such as CD146, CD34, STRO-1 antigen, Thy-1, nestin, Sca-1, and CD133 (25, 26,28,29,31,38,44). SC isolation has been achieved mainly by following protocols that utilize flow cytometry-based sorting of stem cells (11) or positive and negative selections using magnetic beads tagged with antibodies targeting specific markers on the surface of stem cells (17). The most promising stem cell markers are stem cell antigen (Sca-1, Ly-6A/E) and prominin-1 (CD133, AC133), which were found to be expressed on stem cells in the BM, where the positive cells were shown to have a higher potential to differentiate into multiple tissue lineages (19,21,23,33)).…”

Section: Introductionmentioning

confidence: 99%

“…SC isolation has been achieved tial for tissue repair and gene therapy (3,6,34). The first step in stem cell therapy is to obtain the required amount mainly by following protocols that utilize flow cytometry-based sorting of stem cells (11) or positive and nega-of specific population of stem cells. They may be isolated and cultured in vitro in order to increase their num-tive selections using magnetic beads tagged with anti-bodies targeting specific markers on the surface of stem remove cell aggregates and bone fragments.…”

Section: Introductionmentioning

confidence: 99%

Purification of Mouse Bone Marrow-Derived Stem Cells Promotes Ex Vivo Neuronal Differentiation

et al. 2010

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The main objective of this study is to test the potential of specific populations of mouse bone marrowderived stem cells (BMSCs) to differentiate into the neuronal cell lineage. Bone marrow of 33 mice was aspirated under general anesthesia. The collected marrows were analyzed for cell counts, compositions, and percentages of different stem cell types. We used the Midi MACS magnetic separator to purify specific populations of stem cells from the aspirated bone marrow. Cells were analyzed using flow cytometry. We assessed the presence of stem cell antigen-1 (Sca-1 + ) and prominin-1 + cells in the cellular fraction that was depleted of lineage-committed cells (lineage − ). Both purified and nonpurified cells were cultured ex vivo using specific growth media with factors that drive the cells to differentiate into the neuroglial cell types. Cells were then analyzed by flow cytometry for expression of specific neuronal markers. Our results showed that there was an increase of Sca-1 + and prominin-1 + cells in the lineage − fraction over the unpurified BM. After lineage depletion, the percentages of Sca-1 + and prominin-1 + cells increased from 4.9% and 2.6%, up to 76.1% and 59%, respectively. Unpurified mouse BM differentiated into fibroblasts, whereas Sca-1 + cells were able to generate astrocytes. Interestingly, purified prominin-1 + cells were able to generate neuronal cells. Purification of adult bone marrow-derived stem cells enhances their potentiality for differentiating into specific neuronal cell lineages.

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Sterile and disposable fluidic subsystem suitable for clinical high speed fluorescence‐activated cell sorting

Cited by 12 publications

References 37 publications

International Society for Analytical Cytology Biosafety Standard for Sorting of Unfixed Cells

International Society for Analytical Cytology Biosafety Standard for Sorting of Unfixed Cells

Cell type-specific transcriptome analysis in the early Arabidopsis thaliana embryo

Purification of Mouse Bone Marrow-Derived Stem Cells Promotes Ex Vivo Neuronal Differentiation

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