MesenCult™-ACF Plus Medium

Animal component-free medium for human mesenchymal stem cells

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MesenCult™-ACF Plus Medium Kit

Animal component-free medium for human mesenchymal stem cells

1 Kit
Catalog #05445
349 USD

MesenCult™-ACF Plus Culture Kit

Animal component-free medium and attachment substrate for human mesenchymal stem cells

1 Kit
Catalog #05448
399 USD

Required Products


MesenCult™-ACF Plus Medium is a standardized, animal component-free (ACF) and serum-free medium for the isolation and culture of human mesenchymal stromal cells, also known as mesenchymal stem cells (MSCs). MesenCult™-ACF Plus Medium is optimized for the expansion of MSCs in vitro as well as their enumeration using the colony-forming unit-fibroblast (CFU-F) assay. MesenCult™-ACF Plus Medium supports the isolation and long-term growth of human bone marrow-derived MSCs, and cells maintain robust multi-lineage differentiation potential in vitro.
MesenCult™-ACF Plus Medium must be used in conjunction with Animal Component-Free Cell Attachment Substrate (Component #07130) and Animal Component-Free Cell Dissociation Kit (Catalog #05426), providing a complete, defined ACF culture system. Components of Animal Component-Free Cell Attachment Substrate and Animal Component-Free Cell Dissociation Kit are prescreened and tested for optimal cell adherence when cells are cultured with MesenCult™-ACF Plus Medium.
For animal component-free and optimized cryopreservation, MesenCult™-ACF Freezing Medium (Catalog #05490) is recommended for human MSCs previously cultured in MesenCult™ media, including MesenCult™-ACF Plus. For a complete list of related products, including differentiation media available, visit or contact us at
NOTE: Complete MesenCult™-ACF Plus Medium must be supplemented with L-Glutamine (Catalog #07100).
• Animal component-free formulation improves experimental reproducibility.
• Superior cell expansion compared to serum-containing media.
• Cultured MSCs retain robust expansion and tri-lineage differentiation capacities at early and late passages.
• Supports MSC derivation directly from primary human tissue.
MesenCult™-ACF Plus Medium Kit (Catalog #05445)
• MesenCult™-ACF Plus Medium, 500 mL
• MesenCult™-ACF Plus 500X Supplement, 1 mL
MesenCult™-ACF Plus Culture Kit (Catalog #05448)
• MesenCult™-ACF Plus Medium, 500 mL
• MesenCult™-ACF Plus 500X Supplement, 1 mL
• Animal Component-Free Cell Attachment Substrate, 1 mL
Specialized Media
Cell Type:
Mesenchymal Cells, PSC-Derived; Mesenchymal Stem and Progenitor Cells
Cell Culture; Expansion; Maintenance
Area of Interest:
Extracellular Vesicle Research; Stem Cell Biology
Animal Component-Free; Serum-Free

Scientific Resources

Educational Materials


Product Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Data and Publications


Figure 1. CFU-F Assay of Human BM-Derived MSCs Expanded in MesenCult™-ACF Plus Medium and Commercial Media.
(A) An average of 45 CFU-Fs per million cells were observed when BM mononuclear cells were seeded in MesenCult™-ACF Plus (n = 4). An average of 47 and 25 CFU-Fs per million cells were observed when cells were seeded in Commercial Medium 1 (n = 3) and Medium 2 (n = 4), respectively. Vertical lines indicate Standard Error of Mean (SEM). Representative image of CFU-F colonies expanded in (B) MesenCult™-ACF Plus Medium (9 days of culture), (C) Commercial Medium 1 (10 days of culture) and (D) Commercial Medium 2 (10 days of culture). Commercial Medium 1 and Medium 2 were supplemented with 2.5% human AB serum to derive MSCs from BM, as per their protocols for derivation. No addition of serum is required when using MesenCult™-ACF Plus Medium.

Figure 2. Human BM-Derived MSCs Cultured in MesenCult™-ACF Plus Medium Expand Faster than MSCs Cultured in Commercial Xeno-Free and Serum-Free Media.
(A) A greater number of BM-derived MSCs were generated per passage using MesenCult™-ACF Plus Medium (n=4) compared to Commercial Medium 1 (n=3) and Commercial Medium 2 (n=2). (B) Rates of BM-derived MSC expansion were compared between MesenCult™-ACF Plus Medium, Commercial Medium 1, and Commercial Medium 2. The time required to double the number of MSCs using MesenCult™ -ACF Plus Medium (n=4) was shorter than when MSCs were cultured in Commercial Medium 1 (n=3) and Commercial Medium 2 (n=4). Vertical lines indicate Standard Error of Mean (SEM).

Figure 3. Human BM-Derived MSCs Expanded in MesenCult™-ACF Plus Medium Display Multi-Lineage Differentiation Potential.
(A) Human BM-derived MSCs expanded in MesenCult™-ACF Plus Medium differentiated into (B) adipocytes (Oil Red O staining; passage 5), (C) chondrocytes (Alcian Blue staining; passage 4) and (D) osteoblasts (Alizarin Red S staining; passage 5).

Figure 4. Flow Cytometric Analysis of MSCs Cultured in MesenCult™-ACF Plus Medium.
BM-derived MSCs were cultured and expanded in MesenCult™-ACF Plus Medium. At passage 8 MSCs were stained for mesenchymal surface markers (CD73, CD90, CD105,), pericyte marker (CD146) and hematopoietic marker (CD45). MSCs expressed high levels of CD73, CD90, CD105 and CD146 and lacked expression of CD45.


Theranostics 2019

Characterization and Therapeutic Application of Mesenchymal Stem Cells with Neuromesodermal Origin from Human Pluripotent Stem Cells.

H. Wang et al.


Rationale: Mesenchymal stem cells (MSC) hold great promise in the treatment of various diseases including autoimmune diseases, inflammatory diseases, etc., due to their pleiotropic properties. However, largely incongruent data were obtained from different MSC-based clinical trials, which may be partially due to functional heterogeneity among MSC. Here, we attempt to derive homogeneous mesenchymal stem cells with neuromesodermal origin from human pluripotent stem cells (hPSC) and evaluate their functional properties. Methods: Growth factors and/or small molecules were used for the differentiation of human pluripotent stem cells (hPSC) into neuromesodermal progenitors (NMP), which were then cultured in animal component-free and serum-free induction medium for the derivation and long-term expansion of MSC. The resulted NMP-MSC were detailed characterized by analyzing their surface marker expression, proliferation, migration, multipotency, immunomodulatory activity and global gene expression profile. Moreover, the in vivo therapeutic potential of NMP-MSC was detected in a mouse model of contact hypersensitivity (CHS). Results: We demonstrate that NMP-MSC express posterior HOX genes and exhibit characteristics similar to those of bone marrow MSC (BMSC), and NMP-MSC derived from different hPSC lines show high level of similarity in global gene expression profiles. More importantly, NMP-MSC display much stronger immunomodulatory activity than BMSC in vitro and in vivo, as revealed by decreased inflammatory cell infiltration and diminished production of pro-inflammatory cytokines in inflamed tissue of CHS models. Conclusion: Our results identify NMP as a new source of MSC and suggest that functional and homogeneous NMP-MSC could serve as a candidate for MSC-based therapies.
Stem cells international 2019

Gestational Tissue-Derived Human Mesenchymal Stem Cells Use Distinct Combinations of Bioactive Molecules to Suppress the Proliferation of Human Hepatoblastoma and Colorectal Cancer Cells.

N. Paiboon et al.


Background Cancer has been considered a serious global health problem and a leading cause of morbidity and mortality worldwide. Despite recent advances in cancer therapy, treatments of advance stage cancers are mostly ineffective resulting in poor survival of patients. Recent evidences suggest that multipotent human mesenchymal stem cells (hMSCs) play important roles in growth and metastasis of several cancers by enhancing their engraftment and inducing tumor neovascularization. However, the effect of hMSCs on cancer cells is still controversial because there are also evidences demonstrating that hMSCs inhibited growth and metastasis of some cancers. Methods In this study, we investigated the effects of bioactive molecules released from bone marrow and gestational tissue-derived hMSCs on the proliferation of various human cancer cells, including C3A, HT29, A549, Saos-2, and U251. We also characterized the hMSC-derived factors that inhibit cancer cell proliferation by protein fractionation and mass spectrometry analysis. Results We herein make a direct comparison and show that the effects of hMSCs on cancer cell proliferation and migration depend on both hMSC sources and cancer cell types and cancer-derived bioactive molecules did not affect the cancer suppressive capacity of hMSCs. Moreover, hMSCs use distinct combination of bioactive molecules to suppress the proliferation of human hepatoblastoma and colorectal cancer cells. Using protein fractionation and mass spectrometry analysis, we have identified several novel hMSC-derived factors that might be able to suppress cancer cell proliferation. Conclusion We believe that the procedure developed in this study could be used to discover other therapeutically useful molecules released by various hMSC sources for a future in vivo study.