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MesenCultâ„¢-ACF Plus Culture Kit

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

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MesenCultâ„¢-ACF Plus Culture Kit

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

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

What's Included

  • MesenCultâ„¢-ACF Plus Medium, 500 mL
  • MesenCultâ„¢-ACF Plus 500X Supplement, 1 mL
  • Animal Component-Free Cell Attachment Substrate, 1 mL
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.
  1. L-Glutamine
    L-Glutamine

    Cell culture supplement

Overview

Reduce variability in your human mesenchymal stromal cell (MSC; also known as mesenchymal stem cell) cultures and improve experimental reproducibility by using this animal component-free (ACF) and extracellular vesicle (EV)-free medium. MesenCultâ„¢-ACF Plus Culture Kit is optimized so you can derive human MSCs from multiple sources, such as bone marrow or adipose tissue, without serum.

Compared to serum-containing or EV-depleted serum-containing media, MSCs cultured with this kit expand more efficiently (rate and cumulative cell total) and without compromising function. The cultured cells exhibit characteristic MSC surface marker expression and retain robust expansion rate and trilineage differentiation capacities.

This kit is part of a complete ACF workflow—for deriving, expanding, and cryopreserving MSCs, as well as differentiating human pluripotent stem cells into mesenchymal progenitors—that is optimized for efficient and consistent MSC cultures.

For animal component-free and optimized cryopreservation, MesenCultâ„¢-ACF Freezing Medium is recommended for human MSCs previously cultured in MesenCultâ„¢ media, including MesenCultâ„¢-ACF Plus. For a complete list of related products, including available differentiation media, explore our MSC area of interest page or contact us at techsupport@stemcell.com.

NOTE: Complete MesenCultâ„¢-ACF Plus Medium must be supplemented with L-Glutamine. The medium and supplement required for preparation of the complete medium is also available for purchase, without the substrate, as MesenCultâ„¢-ACF Plus Medium Kit.

is the manufacturer of the rhCollagen component of the cell attachment substrate.

The Product may be used for research purposes only. Please contact º£½ÇÆƽâ°æ for any clinical or commercial application of the Product.
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 Category
Animal Component-Free, Serum-Free

Data Figures

Figure 1. CFU-F Assay of Human BM-Derived MSCs Expanded in MesenCultâ„¢-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 MesenCultâ„¢-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) MesenCultâ„¢-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 MesenCultâ„¢-ACF Plus Medium.

Figure 2. Human BM-Derived MSCs Cultured in MesenCultâ„¢-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 MesenCultâ„¢-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 MesenCultâ„¢-ACF Plus Medium, Commercial Medium 1, and Commercial Medium 2. The time required to double the number of MSCs using MesenCultâ„¢ -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 MesenCultâ„¢-ACF Plus Medium Display Multi-Lineage Differentiation Potential.
(A) Human BM-derived MSCs expanded in MesenCultâ„¢-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 MesenCultâ„¢-ACF Plus Medium.
BM-derived MSCs were cultured and expanded in MesenCultâ„¢-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.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Name
Catalog #
05448
Lot #
All
Language
English
Document Type
Product Name
Catalog #
05448
Lot #
All
Language
English
Document Type
Product Name
Catalog #
05448
Lot #
All
Language
English
Document Type
Product Name
Catalog #
05448
Lot #
All
Language
English

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.

Research Area
Workflow Stages

Resources and Publications

Educational Materials (13)

Brochure
Brochure
Brochure
Extracellular Vesicle Generation from Mesenchymal Stromal Cells Using MesenCultâ„¢-ACF Plus
Technical Bulletin
Extracellular Vesicle Generation from Mesenchymal Stromal Cells Using MesenCultâ„¢-ACF Plus
Mesenchymal Stromal Cell-Derived Exosomes: Biogenesis and Cargoes
Wallchart
Mesenchymal Stromal Cell-Derived Exosomes: Biogenesis and Cargoes
The Identity and Properties of Mesenchymal Stem Cells
Wallchart
The Identity and Properties of Mesenchymal Stem Cells
SnapShot: Adipocyte Life Cycle
Wallchart
SnapShot: Adipocyte Life Cycle
MesenCultâ„¢ for Mesenchymal Stromal Cell Derivation, Culture & Differentiation
1:51
Video
MesenCultâ„¢ for Mesenchymal Stromal Cell Derivation, Culture & Differentiation
Correlating TEER Values with Air-Liquid Interface Culture Morphology
3:02
Video
Correlating TEER Values with Air-Liquid Interface Culture Morphology
The Impact of Ancillary Materials on Your Translational Research
17:18
Webinar
The Impact of Ancillary Materials on Your Translational Research
Tools to Accelerate Your MSC and EV Workflows
30:05
Webinar
Tools to Accelerate Your MSC and EV Workflows
The Secret In Vivo Life of "Mesenchymal Stem Cells"
58:00
Webinar
The Secret In Vivo Life of "Mesenchymal Stem Cells"
Mesenchymal Stromal Cells: Markers, Isolation and Culture, Differentiation, and Therapeutic Potential
Mini Review
Mesenchymal Stromal Cells: Markers, Isolation and Culture, Differentiation, and Therapeutic Potentia...

Publications (11)

Dynamic stimulation promotes functional tissue-like organization of a 3D human lymphoid microenvironment model in vitro D. Barozzi et al. Cell Reports Methods 2025 Jul

Abstract

This work focused on generating a three-dimensional (3D) in vitro dynamic model to study chronic lymphocytic leukemia (CLL) cell dissemination, homing, and mechanisms of therapy resistance. We used a gelatin-based, hard porous biomaterial as a support matrix to develop 3D tissue-like models of the human lymph node and bone marrow, which were matured inside bioreactors under dynamic perfusion of medium. Comparing static and dynamic cultures of these 3D constructs revealed that perfusion promoted a tissue-like internal organization of cells, characterized by the expression of specific functional markers and deposition of an intricate extracellular matrix protein network. Recirculation of CLL cells within the dynamic system led to changes in leukemic cell behavior and in the expression of key markers involved in tumor progression. These findings suggest that the model is well suited for investigating the pathophysiological mechanisms of CLL and potentially other hematological malignancies.
Alternative Ways to Obtain Human Mesenchymal Stem Cells from Embryonic Stem Cells Cells 2024 Sep

Abstract

Differentiation approaches to obtain mesenchymal stem cells (MSCs) have gradually developed over the last few decades. The problem is that different protocols give different MSC types, making further research difficult. Here, we tried three different approaches to differentiate embryonic stem cells (ESCs) from early mesoderm to MSCs using serum-containing or xeno-free differentiation medium and observed differences in the cells’ morphology, doubling rate, ability to form colonies, surface marker analysis, and multilineage differentiation potential of the obtained cell lines. We concluded that the xeno-free medium best fits the criteria of MSCs’ morphology, growth kinetics, and surface marker characterization. In contrast, the serum-containing medium gives better potential for further MSC differentiation into osteogenic, chondrogenic, and adipogenic lineages.
Genome-wide sequencing identified extrachromosomal circular DNA as a transcription factor-binding motif of the senescence genes that govern replicative senescence in human mesenchymal stem cells W. Yang et al. Frontiers in Cellular Neuroscience 2024 Aug

Abstract

Mesenchymal stem cells (MSCs) have long been postulated as an important source cell in regenerative medicine. During subculture expansion, mesenchymal stem cell (MSC) senescence diminishes their multi-differentiation capabilities, leading to a loss of therapeutic potential. Up to date, the extrachromosomal circular DNAs (eccDNAs) have been demonstrated to be involved in senescence but the roles of eccDNAs during MSC. Here we explored eccDNA profiles in human bone marrow MSCs (BM-MSCs). EccDNA and mRNA was purified and sequenced, followed by quantification and functional annotation. Moreover, we mapped our datasets with the downloading enhancer and transcription factor-regulated genes to explore the potential role of eccDNAs. Sequentially, gene annotation analysis revealed that the majority of eccDNA were mapped in the intron regions with limited BM-MSC enhancer overlaps. We discovered that these eccDNA motifs in senescent BMSCs acted as motifs for binding transcription factors (TFs) of senescence-related genes. These findings are highly significant for identifying biomarkers of senescence and therapeutic targets in mesenchymal stem cells (MSCs) for future clinical applications. The potential of eccDNA as a stable therapeutic target for senescence-related disorders warrants further investigation, particularly exploring chemically synthesized eccDNAs as transcription factor regulatory elements to reverse cellular senescence.
View All Publications

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