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In vitro mesenchymal trilineage differentiation and extracellular matrix production by adipose and bone marrow derived adult equine multipotent stromal cells on a collagen scaffold.
Abstract: Directed differentiation of adult multipotent stromal cells (MSC) is critical for effective treatment strategies. This study was designed to evaluate the capability of equine MSC from bone marrow (BMSC) and adipose tissue (ASC) on a type I collagen (COLI) scaffold to undergo chondrogenic, osteogenic and adipogenic differentiation and form extracellular matrix (ECM) in vitro. Following determination of surface antigen expression, MSC were loaded into scaffolds in a perfusion bioreactor and loading efficiency was quantified. Cell-scaffold constructs were assessed after loading and 7, 14 and 21 days of culture in stromal or induction medium. Cell number was determined with DNA content, cell viability and spatial uniformity with confocal laser microscopy and cell phenotype and matrix production with light and scanning electron microscopy and mRNA levels. The MSC were positive for CD29 (>90 %), CD44 (>99 %), and CD105 (>60 %). Loading efficiencies were >70 %. The ASC and BMSC cell numbers on scaffolds were affected by culture in induction medium differently. Viable cells remained uniformly distributed in scaffolds for up to 21 days and could be directed to differentiate or to maintain an MSC phenotype. Micro- and ultrastructure showed lineage-specific cell and ECM changes. Lineage-specific mRNA levels differed between ASC and BMSC with induction and changed with time. Based on these results, equine ASC and BMSC differentiate into chondrogenic, osteogenic and adipogenic lineages and form ECM similarly on COLI scaffolds. The collected data supports the potential for equine MSC-COLI constructs to support diverse equine tissue formation for controlled biological studies.
Publication Date: 2013-07-31 PubMed ID: 23892935PubMed Central: PMC3834181DOI: 10.1007/s12015-013-9456-1Google 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
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The study focuses on the manipulation of adult multipotent stromal cells (MSC) to produce different tissues and extracellular matrix (ECM) in vitro for effective treatments. This research uses both bone marrow (BMSC) and adipose tissue (ASC) sources from horses to achieve chondrogenic, osteogenic and adipogenic differentiation.
Research Methodology
- The initial phase involved checking the surface antigen expression of the stromal cells. It was observed that the MSC were positive for CD29 (>90%), CD44 (>99%), and CD105 (>60%).
- The MSC were then loaded into a type I collagen (COLI) scaffold within a perfusion bioreactor, with loading efficiencies noted to be greater than 70%.
- These cell-scaffold constructs were evaluated over three time intervals of 7, 14 and 21 days, under standard stroma or the specific induction that stimulated the MSC towards different lineages.
- Cell number, viability, uniform distribution over time, phenotype, and matrix production were some of the parameters assessed, using various technical methods such as DNA content, confocal laser microscopy, and light and scanning electron microscopy.
Observations and Results
- Both ASC and BMSC were able to maintain their normal state or be directed into chondrogenic, osteogenic and adipogenic differentiation, albeit with some variance in cell numbers depending on the type of induction medium used.
- Viable cells remained uniformly distributed across the scaffolds until the end of the study period (21 days).
- There were distinctive lineage-specific alterations in both the cells themselves and the ECM, observable on a micro and ultrastructural level.
- Differences in lineage-specific mRNA levels between ASC and BMSC during the time course of the study under different induction conditions were noted. This signifies differences in gene activity for different tissue types.
Conclusion of the Study
- This research provides evidence that equine ASC and BMSC can be effectively differentiated into different lineages and form ECM while housed within a COLI scaffold. These results support the potential for equine MSC-COLI constructs to support diverse equine tissue formation for controlled biological studies.
- It opens up potential avenues in stem cell therapy and tissue regeneration applications in equine medicine.
Cite This Article
APA
Xie L, Zhang N, Marsano A, Vunjak-Novakovic G, Zhang Y, Lopez MJ.
(2013).
In vitro mesenchymal trilineage differentiation and extracellular matrix production by adipose and bone marrow derived adult equine multipotent stromal cells on a collagen scaffold.
Stem Cell Rev Rep, 9(6), 858-872.
https://doi.org/10.1007/s12015-013-9456-1 Publication
Researcher Affiliations
- Laboratory for Equine and Comparative Orthopedic Research, Equine Health Studies Program, Department of Veterinary Clinical Sciences, Louisiana State University, Baton Rouge, LA, 70803, USA.
MeSH Terms
- Adipose Tissue / cytology
- Animals
- Bioreactors
- Bone Marrow Cells / cytology
- Bone Marrow Cells / drug effects
- Bone Marrow Cells / ultrastructure
- Cattle
- Cell Count
- Cell Differentiation / drug effects
- Cell Differentiation / genetics
- Cell Lineage / drug effects
- Cell Survival / drug effects
- Cell Survival / genetics
- Collagen / pharmacology
- DNA / metabolism
- Extracellular Matrix / drug effects
- Extracellular Matrix / metabolism
- Extracellular Matrix / ultrastructure
- Flow Cytometry
- Gene Expression Regulation / drug effects
- Horses
- Immunophenotyping
- Mesenchymal Stem Cells / cytology
- Mesenchymal Stem Cells / drug effects
- Mesenchymal Stem Cells / ultrastructure
- Microscopy, Confocal
- Multipotent Stem Cells / cytology
- Multipotent Stem Cells / drug effects
- Multipotent Stem Cells / ultrastructure
- RNA, Messenger / genetics
- RNA, Messenger / metabolism
- Tissue Scaffolds
Grant Funding
- P41 EB002520 / NIBIB NIH HHS
- R01 DE016525 / NIDCR NIH HHS
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