Expansion of mesenchymal stem cells on fibrinogen-rich protein surfaces derived from blood plasma.
Abstract: Mesenchymal stem cells (MSCs) are present in low density in bone marrow and culture expansion is necessary to obtain sufficient numbers for many proposed therapies. Researchers have characterized MSC growth on tissue culture plastic (TCP), although few studies have explored proliferation on other growth substrates. Using adult equine MSCs, we evaluated proliferation on fibrinogen-rich precipitate (FRP) surfaces created from blood plasma. When seeded at 1 × 10(4) cells/cm(2) and passaged five times over 10 days, MSCs on FRP in medium containing fibroblast growth factor 2 (FGF2) resulted in a ∼2.5-fold increase in cell yield relative to TCP. In FGF2-free medium, FRP stimulated a 10.4-fold increase in cell yield over TCP after 10 days, although control cultures maintained in FGF2 on TCP demonstrated that the stimulatory effect of FRP was not as lasting as that of FGF2. Chondrogenic cultures demonstrated that FRP did not affect differentiation. On TCP, MSCs seeded at 500 cells/cm(2) experienced a 4.6-fold increase in cell yield over cultures seeded at 1 × 10(4) cells/cm(2) following 10 days of expansion. In 500 cells/cm(2) cultures, FRP stimulating a two-fold increase in cell yield over TCP without affecting differentiation. Low-density FRP cultures showed a more even distribution of cells than TCP, suggesting that FRP may accelerate proliferation by reducing contact inhibition that slows proliferation. In addition, FRP appears capable of binding FGF2, as FRP surfaces pre-conditioned with FGF2 supported greater proliferation than FGF2-free cultures. Taken together, these factors indicate that substrates obtained from simple and inexpensive processing of blood enhance MSC proliferation and promote efficient coverage of expansion surfaces.
Copyright © 2010 John Wiley & Sons, Ltd.
Publication Date: 2010-12-10 PubMed ID: 21774083DOI: 10.1002/term.352Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research explores the impact of expanding mesenchymal stem cells on fibrinogen-rich protein surfaces derived from blood plasma and finds that these surfaces can stimulate proliferation and distribute cells more evenly.
Objective
The primary aim of this research was to compare mesenchymal stem cell (MSC) growth on standard tissue culture plastic (TCP) with growth on a new surface made from a blood by-product called fibrinogen-rich precipitate (FRP). This was tested using equine MSCs and aimed to determine any advantages of using these new surfaces for proliferation in proposed stem cell therapy applications.
Methodology
- The MSCs were seeded at a density of 1×104 cells per square centimeter and passaged five times over ten days.
- The researchers also experimented with differing cell densities and fibroblast growth factor 2 (FGF2) enrichment to assess the impact on proliferation.
- The cultures were evaluated for proliferation after ten days and differentiation potential was also assessed for chondrogenesis.
- Results were compared to control cultures grown on TCP, both with and without FGF2 supplementation.
Results
- The research found that seeding MSCs on FRP substrates led to an increase in cell yield as compared to TCP.
- In FGF2-enriched medium, FRP facilitated about a 2.5-fold increase in cell yield compared to TCP.
- In FGF2-free medium, FRP stimulated a significant 10.4-fold increase in cell yield over TCP after ten days.
- However, it was noted that the FRP effect wasn’t as long-lasting as the FGF2 effect seen in control cultures on TCP.
- Differentiation potential for chondrogenesis was not affected by FRP substrates, indicating that proliferation benefits do not come at the expense of differentiation capacity.
- Researchers found evidence that lower seeding densities on FRP led to more even cell distribution and greater proliferation.
- It was suggested that one way FRP may promote proliferation is by reducing contact inhibition, a growth restriction that is typically encountered on TCP surfaces.
- FRP surfaces also showed an ability to bind FGF2, further enhancing their proliferation-promoting capacity.
Conclusion
The study demonstrates the benefits of using FRP surfaces derived from blood plasma to culture MSCs. By improving proliferation and distribution of stem cells, the use of FRP surfaces may enhance the efficiency of stem cell therapies. This cost-effective method could therefore revolutionise the expansion of MSCs in regenerative medicine.
Cite This Article
APA
Kisiday JD, Hale BW, Almodovar JL, Lee CM, Kipper MJ, Wayne McIlwraith C, Frisbie DD.
(2010).
Expansion of mesenchymal stem cells on fibrinogen-rich protein surfaces derived from blood plasma.
J Tissue Eng Regen Med, 5(8), 600-611.
https://doi.org/10.1002/term.352 Publication
Researcher Affiliations
- Department of Clinical Science, Colorado State University, Fort Collins, CO, USA. john.kisiday@colostate.edu
MeSH Terms
- Animals
- Cell Count
- Cell Proliferation / drug effects
- Cells, Cultured
- Collagen Type II / metabolism
- Culture Media, Conditioned / pharmacology
- Fibrinogen / pharmacology
- Fibroblast Growth Factor 2 / pharmacology
- Glycosaminoglycans / metabolism
- Horses
- Immunohistochemistry
- Mesenchymal Stem Cells / cytology
- Mesenchymal Stem Cells / drug effects
- Mesenchymal Stem Cells / metabolism
- Plasma / chemistry
- Surface Properties / drug effects
Citations
This article has been cited 11 times.- Trivanovic D, Volkmann N, Stoeckl M, Tertel T, Rudert M, Giebel B, Herrmann M. Enhancement of Immunosuppressive Activity of Mesenchymal Stromal Cells by Platelet-Derived Factors is Accompanied by Apoptotic Priming. Stem Cell Rev Rep 2023 Apr;19(3):713-733.
- Kearney CM, Khatab S, van Buul GM, Plomp SGM, Korthagen NM, Labberté MC, Goodrich LR, Kisiday JD, Van Weeren PR, van Osch GJVM, Brama PAJ. Treatment Effects of Intra-Articular Allogenic Mesenchymal Stem Cell Secretome in an Equine Model of Joint Inflammation. Front Vet Sci 2022;9:907616.
- Teufelsbauer M, Lang C, Plangger A, Rath B, Moser D, Staud C, Radtke C, Neumayer C, Hamilton G. Effects of metformin on human bone-derived mesenchymal stromal cell-breast cancer cell line interactions. Med Oncol 2022 Feb 12;39(5):54.
- Kisiday JD, Liebig BE, Goodrich LR. Adult ovine chondrocytes in expansion culture adopt progenitor cell properties that are favorable for cartilage tissue engineering. J Orthop Res 2020 Sep;38(9):1996-2005.
- Tangtrongsup S, Kisiday JD. Differential Effects of the Antioxidants N-Acetylcysteine and Pyrrolidine Dithiocarbamate on Mesenchymal Stem Cell Chondrogenesis. Cell Mol Bioeng 2019 Apr;12(2):153-163.
- Kisiday JD, Schwartz JA, Tangtrongsup S, Goodrich LR, Grande DA. Culture Conditions that Support Expansion and Chondrogenesis of Middle-Aged Rat Mesenchymal Stem Cells. Cartilage 2020 Jul;11(3):364-373.
- Yao W, Lay YE, Kot A, Liu R, Zhang H, Chen H, Lam K, Lane NE. Improved Mobilization of Exogenous Mesenchymal Stem Cells to Bone for Fracture Healing and Sex Difference. Stem Cells 2016 Oct;34(10):2587-2600.
- Grissom MJ, Temple-Wong MM, Adams MS, Tom M, Schumacher BL, McIlwraith CW, Goodrich LR, Chu CR, Sah RL. Synovial Fluid Lubricant Properties are Transiently Deficient after Arthroscopic Articular Cartilage Defect Repair with Platelet-Enriched Fibrin Alone and with Mesenchymal Stem Cells. Orthop J Sports Med 2014 Jul;2(7).
- García JR, Singh A, García AJ. High fidelity nanopatterning of proteins onto well-defined surfaces through subtractive contact printing. Methods Cell Biol 2014;119:277-92.
- Anderson JA, Little D, Toth AP, Moorman CT 3rd, Tucker BS, Ciccotti MG, Guilak F. Stem cell therapies for knee cartilage repair: the current status of preclinical and clinical studies. Am J Sports Med 2014 Sep;42(9):2253-61.
- Sayyar B, Dodd M, Wen J, Ma S, Marquez-Curtis L, Janowska-Wieczorek A, Hortelano G. Encapsulation of factor IX-engineered mesenchymal stem cells in fibrinogen-alginate microcapsules enhances their viability and transgene secretion. J Tissue Eng 2012;3(1):2041731412462018.
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