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"Smith, Jay"
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Identification and validation of multiple cell surface markers of clinical-grade adipose-derived mesenchymal stromal cells as novel release criteria for good manufacturing practice-compliant production
Background
Clinical translation of mesenchymal stromal cells (MSCs) necessitates basic characterization of the cell product since variability in biological source and processing of MSCs may impact therapeutic outcomes. Although expression of classical cell surface markers (e.g., CD90, CD73, CD105, and CD44) is used to define MSCs, identification of functionally relevant cell surface markers would provide more robust release criteria and options for quality control. In addition, cell surface expression may distinguish between MSCs from different sources, including bone marrow-derived MSCs and clinical-grade adipose-derived MSCs (AMSCs) grown in human platelet lysate (hPL).
Methods
In this work we utilized quantitative PCR, flow cytometry, and RNA-sequencing to characterize AMSCs grown in hPL and validated non-classical markers in 15 clinical-grade donors.
Results
We characterized the surface marker transcriptome of AMSCs, validated the expression of classical markers, and identified nine non-classical markers (i.e., CD36, CD163, CD271, CD200, CD273, CD274, CD146, CD248, and CD140B) that may potentially discriminate AMSCs from other cell types. More importantly, these markers exhibit variability in cell surface expression among different cell isolates from a diverse cohort of donors, including freshly prepared, previously frozen, or proliferative state AMSCs and may be informative when manufacturing cells.
Conclusions
Our study establishes that clinical-grade AMSCs expanded in hPL represent a homogeneous cell culture population according to classical markers,. Additionally, we validated new biomarkers for further AMSC characterization that may provide novel information guiding the development of new release criteria.
Clinical trials
Use of Autologous Bone Marrow Aspirate Concentrate in Painful Knee Osteoarthritis (BMAC): Clinicaltrials.gov
NCT01931007
. Registered August 26, 2013.
MSC for Occlusive Disease of the Kidney: Clinicaltrials.gov
NCT01840540
. Registered April 23, 2013.
Mesenchymal Stem Cell Therapy in Multiple System Atrophy: Clinicaltrials.gov
NCT02315027
. Registered October 31, 2014.
Efficacy and Safety of Adult Human Mesenchymal Stem Cells to Treat Steroid Refractory Acute Graft Versus Host Disease. Clinicaltrials.gov
NCT00366145
. Registered August 17, 2006.
A Dose-escalation Safety Trial for Intrathecal Autologous Mesenchymal Stem Cell Therapy in Amyotrophic Lateral Sclerosis. Clinicaltrials.gov
NCT01609283
. Registered May 18, 2012.
Journal Article
The fourth courier : a novel
A fast-paced espionage thriller for Alan Furst fans set in post-Cold War Poland. It is 1992 in Warsaw, Poland, and the communist era has just ended. A series of grisly murders suddenly becomes an international case when it's feared that the victims may have been couriers smuggling nuclear material out of the defunct Soviet Union. The FBI sends an agent to help with the investigation. When he learns that a Russian physicist who designed a portable atomic bomb has disappeared, the race is on to find him--and the bomb--before it ends up in the wrong hands.
Book
Cheated
In 2010 allegations of an utterly corrupt academic system for student-athletes emerged from the University of North Carolina at Chapel Hill campus, home of the legendary Tar Heels. As the alma mater of Michael Jordan, Larry Brown, Marion Jones, Lawrence Taylor, Rashad McCants, and many others; winner of forty national championships in six different sports; and a partner in one of the best rivalries in sports, UNC-Chapel Hill is a world-famous colossus of college athletics. In the wake of the Wainstein report, however, the fallout from this scandal-and the continuing spotlight on the failings of college athletics-has made the school ground zero in the debate about how the $16 billion college sports industry operates.
Written by UNC professor of history Jay Smith and UNC athletics department whistleblower Mary Willingham,Cheatedexposes the fraudulent inner workings of this famous university. For decades these internal systems have allowed woefully underprepared basketball and football players to take fake courses and earn devalued degrees from one of the nation's top universities while faculty and administrators looked the other way. In unbiased and carefully sourced detail,Cheatedrecounts the academic fraud in UNC's athletics department, even as university leaders focused on minimizing the damage in order to keep the billion-dollar college sports revenue machine functioning. Smith and Willingham make an impassioned argument that the \"student-athletes\" in these programs are being cheated out of what, after all, is promised them in the first place: a college education.
eBook
Live applications of norbormide-based fluorescent probes in Drosophila melanogaster
In this study we investigated the performance of two norbormide (NRB)-derived fluorescent probes, NRBMC009 (green) and NRBZLW0047 (red), on dissected, living larvae of Drosophila, to verify their potential application in live cell imaging confocal microscopy. To this end, larval tissues were exposed to NRB probes alone or in combination with other commercial dyes or GFP-tagged protein markers. Both probes were rapidly internalized by most tissues (except the central nervous system) allowing each organ in the microscope field to be readily distinguished at low magnification. At the cellular level, the probes showed a very similar distribution (except for fat bodies), defined by loss of signal in the nucleus and plasma membrane, and a preferential localization to endoplasmic reticulum (ER) and mitochondria. They also recognized ER and mitochondrial phenotypes in the skeletal muscles of fruit fly models that had loss of function mutations in the atlastin and mitofusin genes, suggesting NRBMC009 and NRBZLW0047 as potentially useful screening tools for characterizing ER and mitochondria morphological alterations. Feeding of larvae and adult Drosophilae with the NRB-derived dyes led to staining of the gut and its epithelial cells, revealing a potential role in food intake assays. In addition, when flies were exposed to either dye over their entire life cycle no apparent functional or morphological abnormalities were detected. Rapid internalization, a bright signal, a compatibility with other available fluorescent probes and GFP-tagged protein markers, and a lack of toxicity make NRBZLW0047 and, particularly, NRBMC009 highly performing fluorescent probes for live cell microscopy studies and food intake assays in Drosophila.
Journal Article
Identification and Preliminary Structure-Activity Relationship Studies of 1,5-Dihydrobenzoe1,4oxazepin-2(3H)-ones That Induce Differentiation of Acute Myeloid Leukemia Cells In Vitro
Acute myeloid leukemia (AML) is the most aggressive type of blood cancer, and there is a continued need for new treatments that are well tolerated and improve long-term survival rates in patients. Induction of differentiation has emerged as a promising alternative to conventional cytotoxic chemotherapy, but known agents lack efficacy in genetically distinct patient populations. Previously, we established a phenotypic screen to identify small molecules that could stimulate differentiation in a range of AML cell lines. Utilising this strategy, a 1,5-dihydrobenzo[e][1,4]oxazepin-2(3H)-one hit compound was identified. Herein, we report the hit validation in vitro, structure-activity relationship (SAR) studies and the pharmacokinetic profiles for selected compounds.
Journal Article
DREADFUL ENEMIES: THE “BEAST,” THE HYENA, AND NATURAL HISTORY IN THE ENLIGHTENMENT
The hypothesis that the beast of the Gévaudan (an intriguingly mysterious killer that roamed southern France in the 1760s) might be an African hyena was not simply a popular and amusing misconception; it reflected an important dimension of the critical spirit driving eighteenth-century science. By historicizing natural discovery and its motivations, this essay uncovers aspects of Enlightenment natural history—namely an attraction to the unknowable and a desire for uncertainty, both reflected in the fascination with the sublime—that only became more marked as the frontiers of knowledge receded. In doing so, the essay shows the distinctively hybrid character of an Enlightenment mentality that savored both illumination and darkness.
Journal Article