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Equine Mesenchymal Stromal Cells from Different Sources Efficiently Differentiate into Hepatocyte-Like Cells.
Abstract: Adult equine hepatocytes have proven challenging to culture long term in vitro as they rapidly lose their morphology and functionality, thus limiting studies on liver function and response to disease. In this study, we describe for the first time the differentiation of equine mesenchymal stromal cells (MSC) from a variety of sources into functional hepatocyte-like cells (HLC). First, we differentiated equine umbilical cord blood (UCB)-derived MSC into HLC and found that these cells exhibited a distinct polygonal morphology, stored glycogen as visualized by periodic acid Schiff's reagent staining, and were positive for albumin and other hepatocyte-specific genes. Second, we demonstrated that UCB-HLC could be revived following cryopreservation and retained their phenotype for at least 10 days. Third, we differentiated three sets of MSC from bone marrow (BM), adipose tissue (AT), and peripheral blood (PB), matched within the same horse. We achieved a 100% differentiation success rate with BM, 0% with AT, and 66% with PB. An additional set of nine PB-MSC samples resulted in an overall success rate of 42% (n = 12), and age or gender did not seem to have an effect on the success of hepatic differentiation from that source. In a final set of experiments, we evaluated the use of these HLC as tools in different fields of biomedical research like virology, to study viral growth, and toxicology, to study chemicals with hepatic toxicity. Equine HLC were found susceptible for infection with the equine herpesviruses type 1 (EHV-1), -2, and -5, and exhibited a more sensitive dose-dependent response to arsenic toxicity than the commonly used human hepatocellular cell line HepG2. Taken together, these data indicate that equine MSC can be efficiently differentiated into HLC and these equine HLC could be a useful tool for in vitro studies.
Publication Date: 2016-05-18 PubMed ID: 27113698DOI: 10.1089/ten.TEC.2015.0403Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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This research investigates a novel method of culturing adult horse liver cells in a lab environment by transitioning them from a variety of original cell types, resulting in ‘hepatocyte-like cells’. They have shown that the resulting cells maintain liver cell characteristics and can be used for research purposes, including the study of liver response to toxins or viruses.
Overview of Research
- The study explores a new means of creating long-term cultures of liver cells for research purposes. Specifically, these liver, or hepatocyte, cells are derived from horses.
- While previously, these cells would quickly lose their characteristics if cultured outside of their natural environment, the application of a new approach involving mesenchymal stromal cells (MSC) gives these liver-like cells better long-term viability.
- These generated hepatocyte-like cells (HLC) have been cultivated from a variety of sources and are convincingly similar to functioning liver cells, demonstrating such characteristics as holding onto glycogen, a key marker of liver cell function.
Methods and Findings
- The research team experimented with MSC from different sources, including umbilical cord blood (UCB), bone marrow, adipose tissue and peripheral blood. They found variable success rates depending on the source. Bone marrow provided a 100% success rate, peripheral blood was at 66%, while adipose tissue didn’t produce any viable HLCs.
- Additionally, they achieved a revival of this type of cells after cryopreservation, sustaining their characteristics for over ten days.
- The researchers also found that horse age and gender had no impact on successful hepatocyte differentiation from these stromal cells.
Applications and Implications
- The research showcases potential uses for these cultured equine HLC in a variety of biomedical research fields, such as virology and toxicology.
- One promising implication is that these HLC can act as an in-vitro representation of hepatocytes for studying liver-specific diseases or toxicities—for instance, they were shown to have a more pronounced response to arsenic poisoning than the human liver cell line typically used in such studies.
- Equine HLC also showed susceptibility to infection by equine herpesviruses, showing promise for virological studies.
- In summary, this method of creating horse liver-like cells from MSCs brings with it opportunities to make significant advancements in a variety of research fields.
Cite This Article
APA
Pennington MR, Curtis TM, Divers TJ, Wagner B, Ness SL, Tennant BC, Van de Walle GR.
(2016).
Equine Mesenchymal Stromal Cells from Different Sources Efficiently Differentiate into Hepatocyte-Like Cells.
Tissue Eng Part C Methods, 22(6), 596-607.
https://doi.org/10.1089/ten.TEC.2015.0403 Publication
Researcher Affiliations
- 1 Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University , Ithaca, New York.
- 1 Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University , Ithaca, New York.
- 2 Department of Biological Sciences, State University of New York at Cortland , Cortland, New York.
- 3 Department of Clinical Sciences, College of Veterinary Medicine, Cornell University , Ithaca, New York.
- 4 Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University , Ithaca, New York.
- 1 Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University , Ithaca, New York.
- 3 Department of Clinical Sciences, College of Veterinary Medicine, Cornell University , Ithaca, New York.
- 3 Department of Clinical Sciences, College of Veterinary Medicine, Cornell University , Ithaca, New York.
- 1 Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University , Ithaca, New York.
MeSH Terms
- Adipose Tissue / cytology
- Animals
- Animals, Newborn
- Bone Marrow Cells / cytology
- Cell Differentiation
- Cells, Cultured
- Female
- Fetal Blood / cytology
- Hep G2 Cells
- Hepatocytes / cytology
- Horses
- Humans
- Male
- Mesenchymal Stem Cells / cytology
- Phenotype
Citations
This article has been cited 6 times.- Harman RM, Churchill KA, Parmar S, Van de Walle GR. Mesenchymal stromal cells isolated from chicken peripheral blood secrete bioactive factors with antimicrobial and regenerative properties. Front Vet Sci 2022;9:949836.
- Harman RM, Patel RS, Fan JC, Park JE, Rosenberg BR, Van de Walle GR. Single-cell RNA sequencing of equine mesenchymal stromal cells from primary donor-matched tissue sources reveals functional heterogeneity in immune modulation and cell motility. Stem Cell Res Ther 2020 Dec 4;11(1):524.
- Wang J, Fu X, Yan Y, Li S, Duan Y, Marie Inglis B, Si W, Zheng B. In vitro differentiation of rhesus macaque bone marrow- and adipose tissue-derived MSCs into hepatocyte-like cells. Exp Ther Med 2020 Jul;20(1):251-260.
- Kamel M, Pavulraj S, Osterrieder K, Azab W. EHV-1 Pathogenesis: Current in vitro Models and Future Perspectives. Front Vet Sci 2019;6:251.
- Harman RM, Yang S, He MK, Van de Walle GR. Antimicrobial peptides secreted by equine mesenchymal stromal cells inhibit the growth of bacteria commonly found in skin wounds. Stem Cell Res Ther 2017 Jul 4;8(1):157.
- Pennington MR, Van de Walle GR. Electric Cell-Substrate Impedance Sensing To Monitor Viral Growth and Study Cellular Responses to Infection with Alphaherpesviruses in Real Time. mSphere 2017 Mar-Apr;2(2).
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