MesenCult™ Proliferation Kit (Human)

Medium for detection of CFU-F and expansion of human mesenchymal stem cells

MesenCult™ Proliferation Kit (Human)

Medium for detection of CFU-F and expansion of human mesenchymal stem cells

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Medium for detection of CFU-F and expansion of human mesenchymal stem cells
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What's Included

  • MesenCult™ MSC Basal Medium (Human), 450 mL (Catalog #05401)
  • MesenCult™ MSC Stimulatory Supplement (Human), 50 mL (Catalog #05402)

Overview

MesenCult™ Proliferation Kit (Human) is a standardized, serum-containing medium for the culture of human mesenchymal stem cells (MSCs). MesenCult™ Proliferation Kit (Human) is optimized for the expansion of human MSCs in vitro as well as for the detection and enumeration of colony-forming unit - fibroblasts (CFU-F). MesenCult™ Proliferation Kit (Human) includes MesenCult™ MSC Basal Medium (Human; 450 mL) and MesenCult™ MSC Stimulatory Supplement (Human; 50 mL).
Subtype
Specialized Media
Cell Type
Mesenchymal Stem and Progenitor Cells
Species
Human
Application
Cell Culture, Colony Assay, Expansion
Brand
MesenCult
Area of Interest
Stem Cell Biology

Data Figures

Figure 1. Human Mesenchymal Stem and Progenitor cells (MSCs) Derived and Culture-Expanded Using the MesenCult™ Proliferation Kit

(A) Human bone marrow mesenchymal stem and progenitor cells (BM MSCs) were derived and culture-expanded for 8 passages in complete MesenCult™ Proliferation Medium (n = 6) and two commercially available media (Commercial Medium 1 and Commercial Medium 2; n = 3). (B) The average doubling time in days over 8 passages of human BM MSCs culture-expanded in complete MesenCult™ Proliferation Medium, and Commercial Medium 1 and 2. Vertical lines indicate standard error of the mean.

Figure 2. Human MSCs Culture-Expanded in Complete MesenCult™ Proliferation Medium Retain Strong Multi-Lineage Differentiation Potential

(A) Human BM-derived MSCs cultured in complete MesenCult™ Proliferation Medium differentiate into (B) adipocytes (Oil Red O staining), (C) chondrocytes (Alcian Blue staining) and (D) osteoblasts (alkaline phosphatase and von Kossa staining).

Figure 3. Flow Cytometric Analysis of Culture-Expanded Human MSCs in Complete MesenCult™ Proliferation Medium

Human BM-derived MSCs from passage 5 were stained for mesenchymal cell surface markers CD73, CD90 and CD105, and the hematopoietic markers CD34 and CD45 (shown in grey shaded areas). BM-derived MSCs are positive for MSC markers and negative for hematopoietic markers. High expression of the endothelial marker, CD146, was also observed in BM-derived MSCs (shown in grey shaded areas). Isotype control is shown by red line.

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
Catalog #
05411
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
05411
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
05411
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.

Resources and Publications

Publications (39)

Thymosin $\beta$4-Enhancing Therapeutic Efficacy of Human Adipose-Derived Stem Cells in Mouse Ischemic Hindlimb Model. J.-H. Kim et al. International journal of molecular sciences 2020 mar

Abstract

Thymosin $\beta$4 (T$\beta$4) is a G-actin sequestering protein that contributes to diverse cellular activities, such as migration and angiogenesis. In this study, the beneficial effects of combined cell therapy with T$\beta$4 and human adipose-derived stem cells (hASCs) in a mouse ischemic hindlimb model were investigated. We observed that exogenous treatment with T$\beta$4 enhanced endogenous TMSB4X mRNA expression and promoted morphological changes (increased cell length) in hASCs. Interestingly, T$\beta$4 induced the active state of hASCs by up-regulating intracellular signaling pathways including the PI3K/AKT/mTOR and MAPK/ERK pathways. Treatment with T$\beta$4 significantly increased cell migration and sprouting from microbeads. Moreover, additional treatment with T$\beta$4 promoted the endothelial differentiation potential of hASCs by up-regulating various angiogenic genes. To evaluate the in vivo effects of the T$\beta$4-hASCs combination on vessel recruitment, dorsal window chambers were transplanted, and the co-treated mice were found to have a significantly increased number of microvessel branches. Transplantation of hASCs in combination with T$\beta$4 was found to improve blood flow and attenuate limb or foot loss post-ischemia compared to transplantation with hASCs alone. Taken together, the therapeutic application of hASCs combined with T$\beta$4 could be effective in enhancing endothelial differentiation and vascularization for treating hindlimb ischemia.
New approach to isolate mesenchymal stem cell (MSC) from human umbilical cord blood. Hussain I et al. Cell biology international 2012 JUL

Abstract

HUCB (human umbilical cord blood) has been frequently used in clinical allogeneic HSC (haemopoietic stem cell) transplant. However, HUCB is poorly recognized as a rich source of MSC (mesenchymal stem cell). The aim of this study has been to establish a new method for isolating large number of MSC from HUCB to recognize it as a good source of MSC. HUCB samples were collected from women following their elective caesarean section. The new method (Clot Spot method) was carried out by explanting HUCB samples in mesencult complete medium and maintained in 37°C, in a 5% CO2 and air incubator. MSC presence was established by quantitative and qualitative immunophenotyping of cells and using FITC attached to MSC phenotypic markers (CD29, CD73, CD44 and CD105). Haematopoietic antibodies (CD34 and CD45) were used as negative control. MSC differentiation was examined in neurogenic and adipogenic media. Immunocytochemistry was carried out for the embryonic markers: SOX2 (sex determining region Y-box 2), OLIG-4 (oligodendrocyte-4) and FABP-4 (fatty acid binding protein-4). The new method was compared with the conventional Rosset Sep method. MSC cultures using the Clot Spot method showed 3-fold increase in proliferation rate compared with conventional method. Also, the cells showed high expression of MSC markers CD29, CD73, CD44 and CD105, but lacked the expression of specific HSC markers (CD34 and CD45). The isolated MSC showed some differentiation by expressing the neurogenic (SOX2 and Olig4) and adipogenic (FABP-4) markers respectively. In conclusion, HUCB is a good source of MSC using this new technique.
Chondrogenesis by chemotactic homing of synovium, bone marrow, and adipose stem cells in vitro. Mendelson A et al. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2011 OCT

Abstract

Cell transplantation has been well explored for cartilage regeneration. We recently showed that the entire articular surface of a synovial joint can regenerate by endogenous cell homing and without cell transplantation. However, the sources of endogenous cells that regenerate articular cartilage remain elusive. Here, we studied whether cytokines not only chemotactically recruit adipose stem cells (ASCs), mesenchymal stem cells (MSCs), and synovium stem cells (SSCs) but also induce chondrogenesis of the recruited cells. Recombinant human transforming growth factor-β3 (TGF-β3; 100 ng) and/or recombinant human stromal derived factor-1β (SDF-1β; 100 ng) was control released into an acellular collagen sponge cube with underlying ASCs, MSCs, or SSCs in monolayer culture. Although all cell types randomly migrated into the acellular collagen sponge cube, TGF-β3 and/or SDF-1β recruited significantly more cells than the cytokine-free control group. In 6 wk, TGF-β3 alone recruited substantial numbers of ASCs (558±65) and MSCs (302±52), whereas codelivery of TGF-β3 and SDF-1β was particularly chemotactic to SSCs (400±120). Proliferation of the recruited cells accounted for some, but far from all, of the observed cellularity. TGF-β3 and SDF-1β codelivery induced significantly higher aggrecan gene expression than the cytokine-free group for ASCs, MSCs, and SSCs. Type II collagen gene expression was also significantly higher for ASCs and SSCs by SDF-1 and TGF-β3 codelivery. Remarkably, the expression of aggrecan and type II collagen was detected among all cell types. Thus, homing of multiple stem/progenitor cell populations may potentially serve as an alternative or adjunctive approach to cell transplantation for cartilage regeneration.