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

Overview

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 www.stemcell.com or contact us at techsupport@stemcell.com.
NOTE: Complete MesenCult™-ACF Plus Medium must be supplemented with L-Glutamine (Catalog #07100).
Advantages:
• 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.
Components:
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
Subtype:
Specialized Media
Cell Type:
Mesenchymal Cells, PSC-Derived; Mesenchymal Stem and Progenitor Cells
Species:
Human
Application:
Cell Culture; Expansion; Maintenance
Brand:
MesenCult
Area of Interest:
Extracellular Vesicle Research; Stem Cell Biology
Formulation:
Animal Component-Free; Serum-Free

Scientific Resources

Educational Materials

(10)

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

Data

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.

Publications

(5)
Blood advances 2019 nov

Human models of NUP98-KDM5A megakaryocytic leukemia in mice contribute to uncovering new biomarkers and therapeutic vulnerabilities.

S. Cardin et al.

Abstract

Acute megakaryoblastic leukemia (AMKL) represents ∼10{\%} of pediatric acute myeloid leukemia cases and typically affects young children ({\textless}3 years of age). It remains plagued with extremely poor treatment outcomes ({\textless}40{\%} cure rates), mostly due to primary chemotherapy refractory disease and/or early relapse. Recurrent and mutually exclusive chimeric fusion oncogenes have been detected in 60{\%} to 70{\%} of cases and include nucleoporin 98 (NUP98) gene rearrangements, most commonly NUP98-KDM5A. Human models of NUP98-KDM5A-driven AMKL capable of faithfully recapitulating the disease have been lacking, and patient samples are rare, further limiting biomarkers and drug discovery. To overcome these impediments, we overexpressed NUP98-KDM5A in human cord blood hematopoietic stem and progenitor cells using a lentiviral-based approach to create physiopathologically relevant disease models. The NUP98-KDM5A fusion oncogene was a potent inducer of maturation arrest, sustaining long-term proliferative and progenitor capacities of engineered cells in optimized culture conditions. Adoptive transfer of NUP98-KDM5A-transformed cells into immunodeficient mice led to multiple subtypes of leukemia, including AMKL, that phenocopy human disease phenotypically and molecularly. The integrative molecular characterization of synthetic and patient NUP98-KDM5A AMKL samples revealed SELP, MPIG6B, and NEO1 as distinctive and novel disease biomarkers. Transcriptomic and proteomic analyses pointed to upregulation of the JAK-STAT signaling pathway in the model AMKL. Both synthetic models and patient-derived xenografts of NUP98-rearranged AMKL showed in vitro therapeutic vulnerability to ruxolitinib, a clinically approved JAK2 inhibitor. Overall, synthetic human AMKL models contribute to defining functional dependencies of rare genotypes of high-fatality pediatric leukemia, which lack effective and rationally designed treatments.
Leukemia 2019 nov

Despite mutation acquisition in hematopoietic stem cells, JMML-propagating cells are not always restricted to this compartment.

A. Caye et al.

Abstract

Juvenile myelomonocytic leukemia (JMML) is a rare aggressive myelodysplastic/myeloproliferative neoplasm of early childhood, initiated by RAS-activating mutations. Genomic analyses have recently described JMML mutational landscape; however, the nature of JMML-propagating cells (JMML-PCs) and the clonal architecture of the disease remained until now elusive. Combining genomic (exome, RNA-seq), Colony forming assay and xenograft studies, we detect the presence of JMML-PCs that faithfully reproduce JMML features including the complex/nonlinear organization of dominant/minor clones, both at diagnosis and relapse. Further integrated analysis also reveals that although the mutations are acquired in hematopoietic stem cells, JMML-PCs are not always restricted to this compartment, highlighting the heterogeneity of the disease during the initiation steps. We show that the hematopoietic stem/progenitor cell phenotype is globally maintained in JMML despite overexpression of CD90/THY-1 in a subset of patients. This study shed new lights into the ontogeny of JMML, and the identity of JMML-PCs, and provides robust models to monitor the disease and test novel therapeutic approaches.
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.

Abstract

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.
Frontiers in pharmacology 2019

Glycyrrhetinic Acid Maintains Intestinal Homeostasis via HuR.

G. Chen et al.

Abstract

Glycyrrhetinic acid (GA) is one of the main components of the traditional Chinese medicine of licorice, which can coordinate and promote the effects of other medicines in the traditional prescription. We found that GA could promote the proliferation, decrease the apoptotic rate, and attenuate DFMO-elicited growth arrest and delay in restitution after wounding in IEC-6 cells via HuR. GA failed to promote proliferation and to suppress apoptosis after silencing HuR by siRNA in IEC-6 cells. Furthermore, with the model of small intestinal organoids developed from intestinal crypt stem cells, we found that GA could increase HuR and its downstream ki67 levels to promote intestinal organoid development. In the in vivo assay, GA was shown to maintain the integrity of the intestinal epithelium under the circumstance of 48 h-fasting in rats via raising HuR and its downstream genes such as EGF, EGFR, and MEK. These results suggested that via HuR modulation, GA could promote intestinal epithelium homeostasis, and therefore contribute to the absorption of constituents from other medicines co-existing in the traditional prescription with licorice in the small intestine. Our results provide a new perspective for understanding the effect of licorice on enhancing the therapeutic effect of traditional prescriptions according to the traditional Chinese medicine theory.
Theranostics 2019

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

H. Wang et al.

Abstract

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.
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