Mesenchymal stem cells (MSCs) have emerged as a powerful tool in regenerative medicine and cell therapy, offering immense potential for treating various diseases and injuries.
A critical factor in realizing this potential is the selection of an appropriate cell culture medium, which plays a vital role in maintaining MSC viability, proliferation, and multipotency.
This article highlights five of the most effective types of media for MSC culture, each offering unique advantages for different research applications and clinical needs.
- Xeno-Free Media: These media, such as StemXVivo® Xeno-Free Human Mesenchymal Stem Cell Expansion Media, are designed to eliminate animal-derived components, reducing the risk of xenogeneic contamination and improving safety for clinical applications.
- Serum-Free Media: Standardized, xeno-free bioprocess medium for the rapid expansion of hMSCs media like RoosterNourish™-MSC-XF offer consistent performance and eliminate the variabiity associated with serum-containing media.
- Specialized MSC Growth Media: Products like PromoCell's Mesenchymal Stem Cell Growth Medium XF are specifically formulated to support robust MSC proliferation while maintaining their multipotent characteristics.
- Traditional FBS-Containing Media: Despite the move towards xeno-free options, some researchers still use fetal bovine serum (FBS) supplemented media for certain applications due to its rich growth factor content.
- Chemically Defined Media: These media, often custom-formulated by leading biotechnology companies, provide the most controlled environment for MSC culture, allowing for precise manipulation of cell behavior and characteristics.
Each of these media types offers distinct advantages and may be preferred depending on the specific research goals, regulatory requirements, and intended applications of the cultured MSCs. As the field of regenerative medicine continues to advance, the choice of culture medium becomes increasingly critical in ensuring the production of high-quality, clinically relevant MSCs.
What is Mesenchymal Stem Cell Medium?
Mesenchymal stem cell medium is a type of cell culture medium used to grow and maintain mesenchymal stem cells (MSCs) in vitro (the lab).
MSCs are a type of adult stem cell that can differentiate into a variety of cell types, including bone, cartilage, and fat cells.
MSCs are often isolated from various tissues, such as bone marrow, adipose tissue, and umbilical cord tissue, and then expanded in culture using a specialized medium.
The composition of mesenchymal stem cell medium can vary depending on the specific application and the source of the MSCs. However, some common components of MSC medium include:
- Basal medium: This is a nutrient-rich solution that provides the basic components necessary for cell growth, such as amino acids, vitamins, and minerals.
- Serum: Fetal bovine serum (FBS) is commonly used as a supplement to provide growth factors and other nutrients that support cell growth and proliferation.
- Antibiotics: Antibiotics such as penicillin and streptomycin are often added to the medium to prevent bacterial contamination.
- Growth factors: Various growth factors, such as fibroblast growth factor (FGF) and transforming growth factor-beta (TGF-β), can be added to the medium to promote MSC proliferation and differentiation.
Overall, mesenchymal stem cell medium is a crucial component of MSC culture and plays a key role in maintaining the stemness and differentiation potential of these cells in vitro.
Mesenchymal stem cell medium is like a special "soup" made in the lab to help a certain kind of stem cell, called mesenchymal stem cells, grow and multiply. The "soup" has all the nutrients and special ingredients these cells need to stay healthy and do their job. It's really important for scientists who are studying how to use these cells to help treat diseases.
Current Trends with Stem Cell Media
The trend in creating clinical products using mesenchymal stem cells (MSCs) is to use xeno-free medium.
Xeno-free medium is a type of culture medium that does not contain any animal-derived components, such as fetal bovine serum (FBS), which is commonly used in traditional MSC culture medium.
The use of xeno-free medium is important for meeting regulatory requirements for clinical-grade MSCs and for reducing the risk of contamination and immune reactions in patients[1].
Xeno-Free Media
One example of a xeno-free medium is OxiumTMEXO, which was evaluated in a study for its ability to sustain increased production of small extracellular vesicles (sEVs) from MSCs.
The study found that OxiumTMEXO displayed a three-fold increase in sEV secretion compared to standard DMEM and a commercially available culture medium developed specifically for sEV production. The higher yield obtained was consistent with several harvest time points.
Another study developed a closed and semi-automated process for passaging and harvesting Wharton's jelly (WJ)-derived MSCs (WJ-MSCs) from multi-layered flasks using counterflow centrifugation.
The WJ-MSCs were expanded using regulatory compliant serum-free xeno-free (SFM XF) medium, which showed comparable cell proliferation and morphology to WJ-MSCs expanded in classic serum-containing media.
The use of xeno-free medium in creating clinical products using MSCs is becoming increasingly popular due to its regulatory compliance, reduced risk of contamination and immune reactions, and comparable cell proliferation and morphology to traditional serum-containing media.
What Are the Benefits of Using a Serum-, Xeno-, and Blood-Free Medium for Clinical-Grade sEV's Production
Using a serum-, xeno-, and blood-free medium for clinical-grade sEV's production has several benefits, including:
- Regulatory compliance: The use of a serum-, xeno-, and blood-free medium meets the regulatory entity requirements of clinical-grade sEV's production.
- Increased sEV production: A study found that OxiumTMEXO, a regulatory complying medium, displayed a three-fold increase of sEV secretion compared to standard DMEM and a commercially available culture medium developed specifically for sEV production[1].
- Reduced risk of contamination and immune reactions: The use of a serum-, xeno-, and blood-free medium reduces the risk of contamination and immune reactions in patients.
- Comparable cell proliferation and morphology: A study found that human mesenchymal stem cells (hMSCs) expanded using regulatory compliant serum-free xeno-free (SFM XF) medium showed comparable cell proliferation and morphology to hMSCs expanded in classic serum-containing media.
In conclusion, using a serum-, xeno-, and blood-free medium for clinical-grade sEV's production is beneficial for regulatory compliance, increased sEV production, reduced risk of contamination and immune reactions, and comparable cell proliferation and morphology.
Conclusion
Choosing the right mesenchymal stem cell (MSC) medium is crucial for successful cell culture and expansion. Based on the latest research and industry trends, here are the top 5 types of media to consider for MSC culture:
- Xeno-Free Media: Products like StemXVivo® Xeno-Free Human MSC Expansion Media eliminate animal-derived components, reducing contamination risks and improving safety for clinical applications.
- Serum-Free Media: Fully defined media such as RoosterNourish™-MSC-XF offer consistent performance and eliminate variability associated with serum-containing media.
- Specialized MSC Growth Media: PromoCell's Mesenchymal Stem Cell Growth Medium XF is specifically formulated to support robust MSC proliferation while maintaining multipotency.
- Chemically Defined Media: These provide the most controlled environment for MSC culture, allowing precise manipulation of cell behavior and characteristics.
- Traditional FBS-Containing Media: While less ideal for clinical applications, some researchers still use fetal bovine serum supplemented media for certain research purposes.
When selecting a medium, consider your specific research goals, regulatory requirements, and intended applications. Xeno-free and serum-free options are increasingly preferred for clinical-grade MSC production due to their safety profile and consistent performance.For optimal results, look for media that support:
- Rapid cell proliferation
- Maintenance of MSC characteristics and multipotency
- Low cellular senescence
- High genetic stability
- Scalability for large-scale production
By choosing the appropriate medium, you can ensure the production of high-quality, clinically relevant MSCs for your research or therapeutic applications.
References
(1) Lo Sicco C, Reverberi D, Pascucci L, Tasso R. A Method for Isolating and Characterizing Mesenchymal Stromal Cell-derived Extracellular Vesicles. Curr Protoc Stem Cell Biol. 2018 Aug;46(1):e55. doi: 10.1002/cpsc.55. Epub 2018 Jun 11. PMID: 29927086.
(2) Yao ZY, Feng BW, Liu CS, Liu YM, Zhou HY, Zhang XH, Jian MQ, Mo JL, Liang YJ, Chen L, Liu XQ, Chen YL, Zhang ZS, He SR. The Application of a Bone Marrow Mesenchymal Stem Cell Membrane in the Vascularization of a Decellularized Tracheal Scaffold. Stem Cells Int. 2021 Mar 5;2021:6624265. doi: 10.1155/2021/6624265. PMID: 33747094; PMCID: PMC7960062.
(3) Figueroa-Valdés AI, de la Fuente C, Hidalgo Y, Vega-Letter AM, Tapia-Limonchi R, Khoury M, Alcayaga-Miranda F. A Chemically Defined, Xeno- and Blood-Free Culture Medium Sustains Increased Production of Small Extracellular Vesicles From Mesenchymal Stem Cells. Front Bioeng Biotechnol. 2021 May 26;9:619930. doi: 10.3389/fbioe.2021.619930. PMID: 34124014; PMCID: PMC8187876.
(4) Figueroa-Valdés AI, de la Fuente C, Hidalgo Y, Vega-Letter AM, Tapia-Limonchi R, Khoury M, Alcayaga-Miranda F. A Chemically Defined, Xeno- and Blood-Free Culture Medium Sustains Increased Production of Small Extracellular Vesicles From Mesenchymal Stem Cells. Front Bioeng Biotechnol. 2021 May 26;9:619930. doi: 10.3389/fbioe.2021.619930. PMID: 34124014; PMCID: PMC8187876.
(5) Lam ATL, Jayaraman P, Becker A, Lim R, Teo KL, Ng J, Oh S. Human Mesenchymal Stem Cell Processing for Clinical Applications Using a Closed Semi-Automated Workflow. J Vis Exp. 2023 Mar 17;(193). doi: 10.3791/64707. PMID: 37010295.
