Full Karyotype Interphase Cell Analysis

Baumgartner, Adi; Hartshorne, Christy; Polyzos, Aris; Weier, Jingly F.; O’Brien, Benjamin

doi:10.1369/0022155418771613

Cited by 5 publications

(10 citation statements)

References 71 publications

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“…Karyotype analysis showed that iBCF displayed the normal camel chromosome number (2n = 74) and structure as pBCF ( Figure 7 ), consistent with previous reports [ 32 ]. Karyotyping is a process of preparing, arranging, and categorizing chromosomes using a cell culture technique in order to encounter various chromosomal abnormalities [ 33 ]. Together with the proliferation test, our study demonstrated that the introduction of telomerase promotes the immortalization of primary BCF, and enhances cell viability and proliferation activity; however, no evidence of any malignant phenotype was associated with the higher growth rate of iBCF.…”

Section: Discussionmentioning

confidence: 99%

Construction and Characterization of Immortalized Fibroblast Cell Line from Bactrian Camel

Yan

Fang

et al. 2023

Life

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Immortalized cell lines with many advantages are widely used in various experimental contexts by many different labs. However, the absence of available cell lines poses difficulties for research in some species, such as camels. To establish an immortalized Bactrian camel fibroblast (iBCF) cell line and understand its biological characteristics, primary fibroblast cells from Bactrian camels were isolated and purified using enzymatic digestion in this study, and telomerase reverse transcriptase (hTERT) vectors were introduced into primary BCF (pBCF) for continuous passage to 80 generations after screening with G418. The cell morphology of different generations was examined under a microscope. Cell cycle and viability were evaluated by flow cytometry and CCK-8 assay, respectively. Cellular genes expression was monitored by qPCR, immunofluorescence, and Western blot, respectively. Chromosomes were determined by karyotyping. The results showed that like most other cells, both pBCF and iBCF were sensitive to nutrient concentrations and adapted to culture in the medium with 4.5 g/L glucose and 10% fetal bovine serum (FBS) concentration. hTERT gene was introduced and stably expressed in iBCF cells, which promoted BCF cell immortalization. The fibroblast specific marker vimentin (VIM) is expressed in both pBCF and iBCF, but epithelial marker cytokeratin18 (CK18) expression is weak in BCF cells. Proliferation and viability detection showed that hTERT-induced iBCF exhibits faster growth rates and higher viability than pBCF. Karyotyping showed that iBCF maintained the same number and morphology of chromosomes as the pBCF. This study demonstrated that we have successfully constructed an immortalized Bactrian camel fibroblast cell line, which was named BCF23. The establishment of the BCF23 cell line provides a foundation for expanding camel-related research.

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Section: Discussionmentioning

confidence: 99%

Construction and Characterization of Immortalized Fibroblast Cell Line from Bactrian Camel

Yan

Fang

et al. 2023

Life

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“…6 However this quantification is not limited to immunohistochemistry and includes DNA or RNA in situ hybridization, or even karyotypic analysis. [70][71][72] These technologies have allowed for increased accuracy and standardization of the biomarker quantification. 73 In recognition of these emerging digital pathology systems and their clinical use, guidelines specific to HER2 immunohistochemistry quantitative image analysis have been published by the CAP.…”

Section: Image Analysis For Biomarkersmentioning

confidence: 99%

“…The FDA has already authorized several products using motorized microscopes and digital cameras or using whole slide images for various biomarkers including ER, PR, HER2, Ki‐67, and p53 6 . However this quantification is not limited to immunohistochemistry and includes DNA or RNA in situ hybridization, or even karyotypic analysis 70–72 . These technologies have allowed for increased accuracy and standardization of the biomarker quantification 73 .…”

Section: Image Analysis and Machine Learning In Pathologymentioning

confidence: 99%

Digital pathology systems enabling quality patient care

Hanna

Ardon

2023

Genes Chromosomes & Cancer

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Pathology laboratories are undergoing digital transformations, adopting innovative technologies to enhance patient care. Digital pathology systems impact clinical, education, and research use cases where pathologists use digital technologies to perform tasks in lieu of using glass slides and a microscope. Pathology professional societies have established clinical validation guidelines, and the US Food and Drug Administration have also authorized digital pathology systems for primary diagnosis, including image analysis and machine learning systems. Whole slide images, or digital slides, can be viewed and navigated similar to glass slides on a microscope. These modern tools not only enable pathologists to practice their routine clinical activities, but can potentially enable digital computational discovery. Assimilation of whole slide images in pathology clinical workflow can further empower machine learning systems to support computer assisted diagnostics. The potential enrichment these systems can provide is unprecedented in the field of pathology. With appropriate integration, these clinical decision support systems will allow pathologists to increase the delivery of quality patient care. This review describes the digital pathology transformation process, applicable clinical use cases, incorporation of image analysis and machine learning systems in the clinical workflow, as well as future technologies that may further disrupt pathology modalities to deliver quality patient care.

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“…Karyotype Analysis. Karyotype analysis was carried out as described before [12,13]. Briefly, iNSCs were cultured in NSC medium for 2 days, and then, the cells were incubated with 100 ng/ml colcemid (Life Technologies, United States) for 90 min.…”

Section: Differentiation Of Inscs To Motormentioning

confidence: 99%

Characterization of Human-Induced Neural Stem Cells and Derivatives following Transplantation into the Central Nervous System of a Nonhuman Primate and Rats

Wang

Zheng

et al. 2022

Stem Cells International

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Neural stem cells (NSCs) and derivatives are potential cellular sources to treat neurological diseases. In the current study, we reprogrammed human peripheral blood mononuclear cells into induced NSCs (iNSCs) and inserted GFP gene into the AAVS1 site for graft tracing. Targeted integration of GFP does not affect the proliferation and differentiation capacity of iNSCs. iNSC-GFP can be further differentiated into dopaminergic precursors (DAPs) and motor neuron precursors (MNPs), respectively. iNSCs were engrafted into the motor cortex and iNSC-DAPs into the striatum and substantia nigra (SN) of a nonhuman primate, respectively. The surviving iNSCs could respond to the microenvironment of the cortex and spontaneously differentiate into mature neurons that extended neurites. iNSC-DAPs survived well and matured into DA neurons following transplantation into the striatum and SN. iNSC-MNPs could also survive and turn into motor neurons after being engrafted into the spinal cord of rats. The results suggest that iNSCs and derivatives have a potential to be used for the treatment of neurological diseases.

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Full Karyotype Interphase Cell Analysis

Cited by 5 publications

References 71 publications

Construction and Characterization of Immortalized Fibroblast Cell Line from Bactrian Camel

Construction and Characterization of Immortalized Fibroblast Cell Line from Bactrian Camel

Digital pathology systems enabling quality patient care

Characterization of Human-Induced Neural Stem Cells and Derivatives following Transplantation into the Central Nervous System of a Nonhuman Primate and Rats

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