Research Containing: Cell culture

Advanced Space Experiment Processor (ADSEP)

by Kevin Tyreon 10 March 2017in Life Sciences

The ADvanced Space Experiment Processor (ADSEP) biotechnology facility contains three independent thermal zones, each accommodating one cassette, and an internal computer that controls the internal functions of all three cassettes. Cassettes can house bioseparation methods, diffusion-cell and mixing-cell capabilities, and cell-culturing equipment.

ADSEP is a fully automated multi-use processing facility for interface with SpaceX’s Dragon, Orbital’s Cygnus or the ISS EXPRESS Rack.

Three independent processing modules can be programmed for totally automated operation.
Processing temperature can be independently monitored and controlled in each of three modules.
Accommodates up to three cassettes, each capable of processing biological samples in space.
From half-stepped mode, ADSEP is capable of conducting up to 44 separate experiments in each cassette assembly.
Biological samples are loaded (preflight) into cassettes that provide appropriate levels of containment.
Processing module doors are opened with two thumb screws, allowing cassettes to be installed in, and removed from, each processing module on orbit.
Cassette interfaces with the processing module through blind-mating power/data connector on back side of cassette.

Space Experiment Cassettes

There are several applications for the unique Techshot ADSEP system:

Cell Dynamics

Cell Celturing (CellCult)

Separations

Biphasic Separation (BISEP)
Microencapsulation
Protein Crystal Growth

Fluid Processing

C. elegans studies
Bacteria studies

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Transient gene and microRNA expression profile changes of confluent human fibroblast cells in spaceflight

by cfynanon 22 August 2016in Biology & Biotechnology No comment

Microgravity, or an altered gravity environment different from the 1 g of the Earth, has been shown to influence global gene expression patterns and protein levels in cultured cells. However, most of the reported studies that have been conducted in space or by using simulated microgravity on the ground have focused on the growth or differentiation of these cells. It has not been specifically addressed whether nonproliferating cultured cells will sense the presence of microgravity in space. In an experiment conducted onboard the International Space Station, confluent human fibroblast cells were fixed after being cultured in space for 3 and 14 d, respectively, to investigate changes in gene and microRNA (miRNA) expression profiles in these cells. Results of the experiment showed that on d 3, both the flown and ground cells were still proliferating slowly, as measured by the percentage of Ki-67(+) cells. Gene and miRNA expression data indicated activation of NF-kappaB and other growth-related pathways that involve hepatocyte growth factor and VEGF as well as the down-regulation of the Let-7 miRNA family. On d 14, when the cells were mostly nonproliferating, the gene and miRNA expression profile of the flight sample was indistinguishable from that of the ground sample. Comparison of gene and miRNA expressions in the d 3 samples, with respect to d 14, revealed that most of the changes observed on d 3 were related to cell growth for both the flown and ground cells. Analysis of cytoskeletal changes via immunohistochemistry staining of the cells with antibodies for alpha-tubulin and fibronectin showed no difference between the flown and ground samples. Taken together, our study suggests that in true nondividing human fibroblast cells in culture, microgravity experienced in space has little effect on gene and miRNA expression profiles.-Zhang, Y., Lu, T., Wong, M., Wang, X., Stodieck, L., Karouia, F., Story, M., Wu, H. Transient gene and microRNA expression profile changes of confluent human fibroblast cells in spaceflight.

Microgravity Reduces the Differentiation and Regenerative Potential of Embryonic Stem Cells

by cfynanon 22 August 2016in Biology & Biotechnology No comment

Mechanical unloading in microgravity is thought to induce tissue degeneration by various mechanisms, including inhibition of regenerative stem cell differentiation. To address this hypothesis, we investigated the effects of microgravity on early lineage commitment of mouse embryonic stem cells (mESCs) using the embryoid body (EB) model of tissue differentiation. We found that exposure to microgravity for 15 days inhibits mESC differentiation and expression of terminal germ layer lineage markers in EBs. Additionally, microgravity-unloaded EBs retained stem cell self-renewal markers, suggesting that mechanical loading at Earth’s gravity is required for normal differentiation of mESCs. Finally, cells recovered from microgravity-unloaded EBs and then cultured at Earth’s gravity showed greater stemness, differentiating more readily into contractile cardiomyocyte colonies. These results indicate that mechanical unloading of stem cells in microgravity inhibits their differentiation and preserves stemness, possibly providing a cellular mechanistic basis for the inhibition of tissue regeneration in space and in disuse conditions on earth.

Microgravity potentiates stem cell proliferation while sustaining the capability of differentiation

by cfynanon 9 June 2015in Biology & Biotechnology No comment

A three-dimensional (3D) clinostat is a device for generating multidirectional G force, resulting in an environment with an average of 10(3) G. Here we report that human mesenchymal stem cells (hMSCs) cultured in a 3D-clinostat (group CL) showed marked proliferation (13-fold in a week) compared with cells cultured under normal conditions of 1 G (group C) (4-fold in a week). Flow cytometry revealed a 6-fold increase in the number of hMSCs double-positive for CD44/CD29 or CD90/CD29 in group CL after 7 days in culture, compared with group C. Telomere length remained the same in cells from both groups during culturing. Group C cells showed increasing expression levels of type II collagen and aggrecan over the culture period, whereas group CL cells showed a decrease to undetectable levels. Pellets of hMSCs from each group were explanted into cartilagedefective mice. The transplants from group CL formed hyaline cartilage after 7 days, whereas the transplants from group C formed only noncartilage tissue containing a small number of cells. These results show that hMSCs cultured in a 3D-clinostat possess the strong proliferative characteristic of stem cells and retain their ability to differentiate into hyaline cartilage after transplantation. On the contrary, cells cultured in a 1-G environment do not maintain these features. Simulated microgravity may thus provide an environment to successfully expand stem cell populations in vitro without culture supplements that can adversely affect stem cell-derived transplantations. This method has significant potential for regenerative medicine and developmental biology.

Identification of proteins involved in inhibition of spheroid formation under microgravity

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Many types of cells transit in vitro from a two- to a three-dimensional growth, when they are exposed to microgravity. The underlying mechanisms are not yet understood. Hence, we investigated the impact of microgravity on protein content and growth behavior. For this purpose the human thyroid cancer cells FTC-133 were seeded either in recently developed cell containers that can endure enhanced physical forces and perform media changes and cell harvesting automatically or in T-25 culture flasks. All cells were cultured for 5 days at 1g. Afterwards, a part of the cell containers were flown to the International Space Station, while another part was kept on the ground. T-25 flasks were mounted on and next to a Random Positioning Machine. The cells were cultured for 12 days under the various conditions, before they were fixed with RNAlater. All fixed cultures showed monolayers, but three-dimensional aggregates were not detected. In a subsequent protein analysis, 180 proteins were identified by mass spectrometry. These proteins did not indicate significant differences between cells exposed to microgravity and their 1g controls. However, they suggest that an enhanced production of proteins related to the extracellular matrix could detain the cells from spheroid formation, while profilin-1 is phosphorylated This article is protected by copyright. All rights reserved

The application of low shear modeled microgravity to 3-D cell biology and tissue engineering

by cfynanon 9 June 2015in Biology & Biotechnology No comment

The practice of cell culture has been virtually unchanged for 100 years. Until recently, life scientists have had to content themselves with two-dimensional cell culture technology. Clearly, living creatures are not constructed in two dimensions and thus it has become widely recognized that in vitro culture systems must become three dimensional to correctly model in vivo biology. Attempts to modify conventional 2-D culture technology to accommodate 3-D cell growth such as embedding cells in extracellular matrix have demonstrated the superiority of concept. Nevertheless, there are serious drawbacks to this approach including limited mass transport and lack of scalability. Recently, a new cell culture technology developed at NASA to study the effects of microgravity on cells has emerged to solve many of the problems of 3-D cell culture. The technology, the Rotating Wall Vessel (RWV) is a single axis clinostat consisting of a fluid-filled, cylindrical, horizontally rotating culture vessel. Cells placed in this environment are suspended by the resolution of the gravitational, centrifugal and Coriolis forces with extremely low mechanical shear. These conditions, which have been called “low shear modeled microgravity”, enable cells to assemble into tissue-like aggregates with high mass transport of nutrients, oxygen and wastes. Examples of the use of the RWV for basic cell biology research and tissue engineering applications are discussed.

Human Hematopoietic Progenitor Cells Grow Faster Under Rotational Laminar Flows

by cfynanon 9 June 2015in Biology & Biotechnology No comment

We report significant and reproducible growth acceleration of human progenitor cells when exposed to rotational flow when compared with stationary conditions. Nonenriched CD34+ umbilical cord derived human hematopoietic progenitor cells were cultured in Petri dishes located at different radial distances with respect to the central axis of a rotating platform. Growth dynamics under 3 or 5 rpm agitation was compared against that observed under typical stationary conditions. Cells cultured at 3 or 5 rpm exhibited (a) the absence of a latency phase, (h) an increase in final cell concentrations by 54-58.5%, and (c) reduced doubling time in their exponential phase by 12-16% in comparison with stationary culture. Cells grown under rotational agitation were confirmed to remain CD34+ by PCR. These results document a significant positive effect of exposure to laminar flow fields on the growth of human hematopoietic progenitor cells. (C) 2010 American Institute of Chemical Engineers Biotechnol. Prog. 26: 1465-1473, 2010

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Multi-use Variable-gravity Platform (MVP)

by Mallory Shorton 3 June 2015in Life Sciences

Multi-use Variable-gravity Platform (MVP) – Dual Centrifuges Simultaneously Providing 0-2g Each

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Cell culturing (adherent and suspended)
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Research Containing: Cell culture

Advanced Space Experiment Processor (ADSEP)

Space Experiment Cassettes

Cell Dynamics

Separations

Fluid Processing

Learn more about Techshot

Transient gene and microRNA expression profile changes of confluent human fibroblast cells in spaceflight

Microgravity Reduces the Differentiation and Regenerative Potential of Embryonic Stem Cells

Microgravity potentiates stem cell proliferation while sustaining the capability of differentiation

Identification of proteins involved in inhibition of spheroid formation under microgravity

The application of low shear modeled microgravity to 3-D cell biology and tissue engineering

Human Hematopoietic Progenitor Cells Grow Faster Under Rotational Laminar Flows

Multi-use Variable-gravity Platform (MVP)

Multi-use Variable-gravity Platform (MVP) – Dual Centrifuges Simultaneously Providing 0-2g Each

Learn more about Techshot