This year was challenging to get through in many ways, but the space world was kicking ass. From Blue Origin and SpaceX making vertical booster landings look easy to the introduction of several new launch vehicles and launch sites, it was a busy space year. Overall, there were 85 launches and 3 failures in 2016 including 4 crew and 7 cargo flights to the ISS.
There were several notable firsts for ISS research as well, especially for those interested in microgravity molecular biology. Because of the multi-step, hands-on nature of life science benchwork, significant progress in this field has lumbered along from a lack of development of even the most basic assay resources for the ISS.
The majority of life science projects flown are one or two step experiments performed in self-contained modules with minimal to no crew interaction. The samples or modules is then returned and analyzed post-flight. If it didn’t fail, you may have the money to fly again. Maybe not. These constraints have seriously limited the pace of what can be accomplished in this field.
The developments seen on the ISS this year are important steps forward to remedy these challenges and create a more relevant and productive orbiting national laboratory. In no specific order, I’ve summarized what I think are 5 of the most significant projects that happened on the ISS in 2016. What was your favorite?
Click here for NASA’s list of ISS research.
PI: Aaron Burton, Ph.D., NASA JSC, Houston, TX, United States
Hardware: Oxford Nanopore Technologies MinION, modified for flight in cooperation with NASA. Samples provided by NASA.
Sponsor/Funding: NASA Technology Demonstration Office
The Oxford Nanopore MinION has been available to researchers for several years now and commercially available since 2015. Its small footprint, lack of moving parts and relatively easy to use software made it an easy choice for a first try at DNA sequencing in microgravity. While the MinION itself is currently limited in its capability, this project is certainly a grand start to having an in situ DNA sequencing resource on the ISS.
Previously, one way to check for microbial contamination was for the crew to swab areas of the ISS and then freeze the samples until they were returned to scientists on Earth. With currently only two vehicles having sample return capabilities (Soyuz and Dragon), this is a slow process. If there really was something aggressive and infectious growing up there, it could be months until it was discovered and dealt with. The gravity (haha) of this problem increases if we ever embark on longer, more distant spaceflight missions. It’s the stuff of sci-fi horror movies, actually.
To change this slow process, this project had two goals: to test if DNA sequencing with the MinION can be performed in micro-g and how can the procedures be optimized for future samples? The experiment consisted of analyzing three standard DNA sample libraries of lambda bacteriophage (~48.5 kb, genomic DNA), Escherichia coli (~4.7 Mb, genomic DNA) and Mus musculus (BALB/c) (~16.3 kb mitochondrial DNA) that were prepared on Earth before launch.
One of the biggest worries in sample handling for this investigation was the inclusion of bubbles into the system. Bubbles are a nuisance in a pipette tip in 1g let alone micro-g and the usual flushing techniques used to move them out of tips and tubing only results in frustration in microgravity.
Luckily, astronaut Kate Rubins breezed through the procedures without such problems and most importantly, initial results showed the the MinION performed comparably to one on Earth. The last numbers I saw was that the microgravity MinION had completed 80,000 total reads with an average read length of ~6,000 bp. Not bad at all.
PI: Macarena Parra, NASA Ames Space Biosciences Division and Julie Schonfeld, NASA, Engineering Directorate, NASA’s Ames Research Center
Hardware: Ames Research Center, Moffett Field, CA
Sponsor/Funding: NASA National Lab
After losing the first version of WetLab-2 on the SpaceX-7 vehicle failure last year, it was great to see this excellent contribution to the ISS molecular biology assay collection safely arrive. Another common molecular biology Earth lab assay, WetLab-2 is a qPCR system that combines sample prep ability (cell lysis, RNA purification, etc.), a sample transfer tool and a commercial-off-the-shelf (COTS) 16 sample thermocycler/qPCR device from Cephid called the SmartCycler.
Over several sessions in April, astronaut Jeff Williams used lyophilized DNA, E. coli cells and mouse liver tissue samples that arrived with the device to validate the components and procedures of the system. With only a small hiccup due to bubbles, the sessions went very well with the data being comparable to the Earth system. It’s remarkable that the data from this (and the DNA sequencing) were simply e-mailed from the ISS to the investigators, saving money and time on their projects.
PI: David Scott Copeland, The Boeing Company, Pasadena, TX, United States
Hardware: miniPCR in collaboration with Boeing
Sponsor/Funding: NASA National Lab, CASIS, Boeing, New England BioLabs, Math for America
I wasn’t kidding when I said the ISS made notable strides in upping its research game this year. Want to just perform PCR not qPCR on the ISS? Well now you can do that, too. This project was the winning “Genes in Space” entry from a 16 year old high school student named Anna-Sophia Boguraev.
“Genes in Space” is a yearly contest for students from 7-12th grade that propose a DNA experiment using microgravity and the miniPCR system. In its second year, the contest is a collaboration with miniPCR, the maker of the small thermocycler that was flown to the ISS, Boeing, New England BioLabs, Math for America and CASIS.
Previous data supports that spaceflight can alter the methylation status of genes in rice as well as human lymphocytes in simulated micro-g. Anna-Sophia proposed using PCR to measure the methylation status of in vitro genes involved in the immune response and see if this could be a possible mechanism that leads to a deficient immune response in astronauts.
Using a standard of measuring DNA methylation status called bisulfite conversion, zebrafish DNA samples prepared before launch were placed in the miniPCR and allowed to perform a thermocycler program loaded beforehand. When completed, the samples were removed from the hardware and placed in cold stowage for the return flight and ground analysis. Ground samples were also ran in an Earth laboratory and analysis will compare the two. The runs on the ISS went without any problems and it will be great to hear how the data from the micro-g samples turned out.
The miniPCR thermocycler itself is pretty sweet. I’ve often wondered why thermocyclers (and quite a lot of other lab equipment) are still so expensive and large nowadays since electronics are smaller and relatively cheap. Well, Zeke Alvarez-Saavedra, Ph.D. and Sebastian Kraves, Ph.D. wondered the same thing and decided to see if they could do something about it.
After a successful Kickstarter campaign, an 8 sample thermocycler about ⅕ the size of a normal one (2 x 5 x 4 in., ~1 lb.) was born. It runs with an app on your phone or tablet and has several standard PCR system components available (electrophoresis, transilluminator, etc). Perfect for the ISS. I hope to eventually see the capability to analyze the samples on the ISS as well.
PI: Joseph C. Wu, M.D., Ph.D., Stanford University School of Medicine, Stanford, CA, United States
Hardware: BioServe Space Technologies, University of Colorado, Boulder, CO, United States
Sponsor/Funding: NASA National Lab, CASIS
Microgravity affects astronaut’s cardiovascular system in several ways, including transient tachycardia and arrhythmia, changes in the heart’s shape such as having a more spherical form, decreases in left ventricular mass, and orthostatic intolerance to name a few. Like the majority of physiological changes observed in microgravity, there is little known about the molecular basis of why these changes are occurring. This experiment hopes to shed a little light on the molecular basis of these changes.
This collaboration with Stanford University, The Ohio State University, Brigham and Women’s Hospital, NASA and CASIS used cardiomyocytes from induced pluripotent stem cells (iPSC) to study cellular morphology and genetic changes that occur during exposure to microgravity. It’s important to note that they used iPSC’s from skin samples, since cardiomyocytes are hard to come by (unless you know of anyone willing to give their still beating heart to science) and therefore research in these areas have been hard to perform.
Samples of the cells were cultured for ~30 days using BioServe’s BioCell cartridges, observed with BioServe’s microscope for confluency and overall cell health and then fixed with RNAlAter and placed in cold stowage for return to Earth. Initial results showed the cells were beating arrhythmically while in micro-g, but settled back to a synchronous pulse when returned to Earth.
Astronaut Kate Rubins let out a “that’s so cool” when the cells reached confluency and began beating in unison. I agree. Cool indeed. Keep an eye out for the paper later this year. Should be some interesting results.
PI’s: Gioia D. Massa, Kennedy Space Center, Kennedy Space Center, FL, United States
Robert C. Morrow, Ph.D., Orbital Technologies Corporation, Madison, WI, United States
Raymond M. Wheeler, Kennedy Space Center, Cape Canaveral, FL, United States
Hardware: Orbital Technologies Corp. (ORBITEC), Madison, Wisconsin, KSC
Sponsor/Funding: NASA Human Exploration and Operations Mission Directorate (HEOMD), SBIR
So why is eating lettuce on the ISS a big deal? Because it’s the stuff of science fiction and no large scale plant growth has ever been performed before Veggie (mold and fungus on ISS/Mir doesn’t count). And also because it’s a project that’s been ~30 years in the making.
The Veggie system idea started in the late 80’s at the the University of Wisconsin and with some help from NASA, patents were awarded that evolved into the now ubiquitous red and blue LED lighting system seen on many commercially available plant growth systems. The idea was further developed at Kennedy Space Center through the creation of other plant growth projects for the ISS: Advanced ASTROCULTURE™, Advanced Biological Research System, the Vegetable Production System, and the Plant Habitat.
This wasn’t the first time edible plants had been grown in space though, with super dwarf wheat being grown in Greenhouse-II on Mir in the 1990’s and 17 “Rasteniya” experiments grown in the (much smaller than Veggie) Lada Greenhouse on the Russian side of the ISS from 2002-2011. The U.S. actually collaborated with the Russians on use of Lada from 2008-2010 to help define and hone procedures for growing and eating plants that would be later incorporated in Veggie.
After a few ground test facilities were made, Veg-01 flight hardware design was finalized and launched to the ISS in 2014. An initial demonstration of Outredgeous romaine lettuce and zinnias were grown, but the crew was not officially allowed to consume the lettuce until samples were returned to Earth for contamination analysis. Finally in 2016, the crew was given the all clear from NASA to eat what they had harvested. But only after the cleaned the leaves with a citric acid based sanitizer first. Yum.
After eating, U.S. astronaut Scott Kelly quipped poetically, “That’s one small bite for a man, one giant leaf for mankind.” I think that pun was planted.
PI: Michael P. Snyder, M.S. Aeronautical & Astronautical Engineering, Made in Space, Moffett Field, CA, United States
Hardware: Made In Space
Sponsor/Funding: NASA National Lab/Private
Like the iPhone, sometimes you don’t know what you want until you have it. It’s not just the idea of having a 3D printer on the ISS for making things you need, but the security of being able to make things you don’t know you’ll need.
Made In Space’s new version of their 3D printer arrived on the ISS this year and is now taking orders to print your ideas in microgravity. This version is twice as large as the previous one (also still on the ISS, but not for public use), can print with a variety of materials and has modular, replaceable parts designed to help it last the lifetime of the ISS. And while they’ve been kinda quiet about it, there have been almost 70 items printed on the ISS.
I’ve heard researchers opine that they didn’t get the data they wanted because “only if this-and-this” were on station. So here’s your chance to fix that: I challenge you to come up with a way to utilize a microgravity 3D printed item in your next experiment.