Isolation, Maintenance and Differentiation of Primary Tracheal Basal Cells from Adult Rhesus Macaque

Anna E. Engler; Gustavo Mostoslavsky; Lisa Miller; Jason R. Rock

doi:10.3390/mps2040079

Isolation, Maintenance and Differentiation of Primary Tracheal Basal Cells from Adult Rhesus Macaque

Methods and Protocols ◽

10.3390/mps2040079 ◽

2019 ◽

Vol 2 (4) ◽

pp. 79

Author(s):

Anna E. Engler ◽

Gustavo Mostoslavsky ◽

Lisa Miller ◽

Jason R. Rock

Keyword(s):

Rhesus Macaque ◽

Rhesus Macaques ◽

Enzymatic Digestion ◽

Basal Cells ◽

Early Passage ◽

Long Term Storage ◽

Air Liquid Interface ◽

Nonadherent Cells ◽

Level 2

In this report, we describe methodologies for the isolation and culture of primary rhesus macaque tracheal basal cells, their cryopreservation, long term storage and differentiation. These are comparable to state-of-the-art protocols that have been developed for mouse and human airway basal cells. This method is based on the use of proprietary media, providing an easily reproducible and applicable protocol for usage in biosafety level 2 (BSL2) settings. Tracheas from rhesus macaques were isolated after animal euthanasia and subjected to enzymatic digestion overnight. Cells of the epithelial layer were scraped off of the trachea for cell culture. Twenty-four hours after plating basal cells had attached and nonadherent cells were removed. First passages of basal cells can be frozen for early passage storage in liquid nitrogen or propagated and differentiated on an air–liquid interface and in a tracheosphere assay up to passage seven. This protocol provides a platform for the analysis of basal cells from a close evolutionary relative to humans.

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Fallopian Tube Basal Stem Cells Reproducing the Epithelial Sheets In Vitro—Stem Cell of Fallopian Epithelium

Biomolecules ◽

10.3390/biom10091270 ◽

2020 ◽

Vol 10 (9) ◽

pp. 1270

Author(s):

Maobi Zhu ◽

Tomohiko Iwano ◽

Sen Takeda

Keyword(s):

Stem Cells ◽

Fallopian Tube ◽

Culture Method ◽

Reproductive Organ ◽

Basal Cells ◽

Ciliated Cells ◽

Epithelial Sheets ◽

Air Liquid Interface

The fallopian tube (FT) is an important reproductive organ in females. The luminal epithelium of the FT is composed of highly polarized secretory and ciliated cells. Recently, accumulating lines of evidence have suggested that the origin of high-grade serous ovarian carcinoma (HGSC) is fallopian tube epithelial cells (FTECs). Due to the lack of a high-fidelity model for FTECs in vitro, homeostasis, differentiation, as well as the transformation of FTECs are still enigmatic. In this study, we optimized the culture condition for the stable expansion of basal stem cells, as well as inducing differentiation of basal cells into polarized secretory and ciliated cells in the air–liquid interface (ALI) condition suitable for long-term culture. This storable culture method of FTECs provides a versatile platform for studying differentiation mechanisms, intercellular communication, and transformation to HGSC, as well as the physiological function of the FT in vitro.

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Origins and Long-Term Patterns of Copy-Number Variation in Rhesus Macaques

Molecular Biology and Evolution ◽

10.1093/molbev/msaa303 ◽

2020 ◽

Author(s):

Gregg W C Thomas ◽

Richard J Wang ◽

Jelena Nguyen ◽

R Alan Harris ◽

Muthuswamy Raveendran ◽

...

Keyword(s):

Copy Number Variation ◽

Rhesus Macaque ◽

Copy Number ◽

Rhesus Macaques ◽

Gene Copy ◽

Similar Data ◽

Single Nucleotide Variants ◽

Data Set ◽

Number Variation

Abstract Mutations play a key role in the development of disease in an individual and the evolution of traits within species. Recent work in humans and other primates has clarified the origins and patterns of single-nucleotide variants, showing that most arise in the father’s germline during spermatogenesis. It remains unknown whether larger mutations, such as deletions and duplications of hundreds or thousands of nucleotides, follow similar patterns. Such mutations lead to copy-number variation (CNV) within and between species, and can have profound effects by deleting or duplicating genes. Here, we analyze patterns of CNV mutations in 32 rhesus macaque individuals from 14 parent–offspring trios. We find the rate of CNV mutations per generation is low (less than one per genome) and we observe no correlation between parental age and the number of CNVs that are passed on to offspring. We also examine segregating CNVs within the rhesus macaque sample and compare them to a similar data set from humans, finding that both species have far more segregating deletions than duplications. We contrast this with long-term patterns of gene copy-number evolution between 17 mammals, where the proportion of deletions that become fixed along the macaque lineage is much smaller than the proportion of segregating deletions. These results suggest purifying selection acting on deletions, such that the majority of them are removed from the population over time. Rhesus macaques are an important biomedical model organism, so these results will aid in our understanding of this species and the disease models it supports.

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Origins and long-term patterns of copy-number variation in rhesus macaques

10.1101/749416 ◽

2019 ◽

Author(s):

Gregg W.C Thomas ◽

Richard J. Wang ◽

Jelena Nguyen ◽

R. Alan Harris ◽

Muthuswamy Raveendran ◽

...

Keyword(s):

Copy Number Variation ◽

Rhesus Macaque ◽

Copy Number ◽

Rhesus Macaques ◽

Gene Copy Number ◽

Model Organism ◽

Gene Copy ◽

Single Nucleotide Variants ◽

Number Variation

AbstractMutations play a key role in the development of disease in an individual and the evolution of traits within species. Recent work in humans and other primates has clarified the origins and patterns of single nucleotide variants, showing that most arise in the father’s germline during spermatogenesis. It remains unknown whether larger mutations, such as deletions and duplications of hundreds or thousands of nucleotides, follow similar patterns. Such mutations lead to copy-number variation (CNV) within and between species, and can have profound effects by deleting or duplicating genes. Here we analyze patterns of CNV mutations in 32 rhesus macaque individuals from 14 parent-offspring trios. We find the rate of CNV mutations per generation is low (less than one per genome) and we observe no correlation between parental age and the number of CNVs that are passed on to offspring. We also examine segregating CNVs within the rhesus macaque sample and compare them to a similar dataset from humans, finding that both species have far more segregating deletions than duplications. We contrast this with long-term patterns of gene copy-number evolution between 17 mammals, where the proportion of deletions that become fixed along the macaque lineage is much smaller than the proportion of segregating deletions. These results suggest purifying selection acting on deletions, such that the majority of them are removed from the population over time. Rhesus macaques are an important biomedical model organism, so these results will aid in our understanding of this species and the disease models it supports.

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Preparation And elemental analysis of ancient fibers

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100126846 ◽

1987 ◽

Vol 45 ◽

pp. 410-411

Author(s):

Allen Angel ◽

Kathryn A. Jakes

Keyword(s):

Cross Sections ◽

Physical Structure ◽

Energy Dispersive ◽

Archaeological Sites ◽

X Ray ◽

Long Term Storage ◽

Backscattered Electron ◽

Electron Imaging

Fabrics recovered from archaeological sites often are so badly degraded that fiber identification based on physical morphology is difficult. Although diagenetic changes may be viewed as destructive to factors necessary for the discernment of fiber information, changes occurring during any stage of a fiber's lifetime leave a record within the fiber's chemical and physical structure. These alterations may offer valuable clues to understanding the conditions of the fiber's growth, fiber preparation and fabric processing technology and conditions of burial or long term storage (1).Energy dispersive spectrometry has been reported to be suitable for determination of mordant treatment on historic fibers (2,3) and has been used to characterize metal wrapping of combination yarns (4,5). In this study, a technique is developed which provides fractured cross sections of fibers for x-ray analysis and elemental mapping. In addition, backscattered electron imaging (BSI) and energy dispersive x-ray microanalysis (EDS) are utilized to correlate elements to their distribution in fibers.

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Modelling of Moisture Movement in Wood during Outdoor Storage

Nonlinear Analysis Modelling and Control ◽

10.15388/na.2001.6.1.15210 ◽

2001 ◽

Vol 6 (2) ◽

pp. 3-14 ◽

Cited By ~ 23

Author(s):

R. Baronas ◽

F. Ivanauskas ◽

I. Juodeikienė ◽

A. Kajalavičius

Keyword(s):

Experimental Data ◽

Finite Difference ◽

Climatic Conditions ◽

Two Dimensional ◽

Moisture Movement ◽

Finite Difference Technique ◽

Long Term Storage ◽

Space Formulation ◽

Term Storage

A model of moisture movement in wood is presented in this paper in a two-dimensional-in-space formulation. The finite-difference technique has been used in order to obtain the solution of the problem. The model was applied to predict the moisture content in sawn boards from pine during long term storage under outdoor climatic conditions. The satisfactory agreement between the numerical solution and experimental data was obtained.

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