MesenCult™ Osteogenic Differentiation Kit (Human)

For the in vitro differentiation of human MSCs into osteoblasts

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MesenCult™ Osteogenic Differentiation Kit (Human)

For the in vitro differentiation of human MSCs into osteoblasts

1 Kit
Catalog #05465
264 USD

Overview

MesenCult™ Osteogenic Differentiation Kit (Human) is specifically formulated for the in vitro differentiation of human mesenchymal stem and progenitor cells (MSCs) into cells of the osteogenic lineage. This kit is suitable for the differentiation of human bone marrow (BM)-derived MSCs previously culture-expanded in serum-containing medium (e.g. MesenCult™ Proliferation Kit [Human; Catalog #05411] or MesenCult™-hPL [Human; Catalog #05439]) or animal component-free MesenCult™-ACF Plus Medium [Catalog #05445]).
Advantages:
• Compatible with human MSCs previously culture-expanded in MesenCult™ expansion media.
• Available in an easy-to-use two-component format.
• Rigorous raw material screening and quality control minimize lot-to-lot variability.
Components:
  • MesenCult™ Osteogenic Differentiation Basal Medium (Human), 200 mL
  • MesenCult™ Osteogenic Differentiation 5X Supplement (Human), 50 mL
Cell Type:
Mesenchymal Stem and Progenitor Cells; Osteoblasts
Application:
Cell Culture; Differentiation
Brand:
MesenCult
Area of Interest:
Stem Cell Biology

Scientific Resources

Educational Materials

(5)

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. Robust Bone Marrow Mesenchymal Stem and Progenitor Cells (BM MSCs) Osteogenic Differentiation is Achieved in 14 days

Human BM MSCs were derived and expanded for 3 passages using the MesenCult™-ACF Culture Kit (Catalog #05449), MesenCult™ Proliferation Kit (Catalog #05411) or in MesenCult™-hPL Medium (05439). Once MSCs reach greater than 95% confluency, MesenCult™ Osteogenic medium was added to each MSC culture. Osteogenic differentiation was observed within 14 days of induction as indicated by strong alkaline phosphatase activity (red stain) and bone mineralization by the von Kossa method (black stain). Negative controls of undifferentiated MSC cultures were kept in each MesenCult™ MSC expansion media for the same time period. Negative Controls show little or no alkaline phosphatase activity and bone mineralization.

Figure 2. MesenCult™ Osteogenic Medium Leads to Faster and Stronger Osteogenic Differentiation When Compared to A Commercial Osteogenic Differentiation Medium

BM MSCs derived and expanded in MesenCult™-ACF or MesenCult™ Proliferation medium were differentiated for 14 days in either MesenCult™ Osteogenic or another Commercial Osteogenic Medium. Differentiation assays using the MesenCult™ Osteogenic medium displayed stronger alkaline phosphatase activity (red stain) and bone mineralization (black stain) when compared to cultures differentiated with a Commercial Osteogenic Medium.

Figure 3. Osteogenic Differentiation of ES-Derived Mesenchymal Progenitor Cells (MPCs)

Mesenchymal progenitor cells (MPCs) were derived from a human iPS or ES cell line using the STEMdiff™ Mesenchymal Progenitor Kit (Catalog #05240) and expanded for 18 or 17 passages, respectively. Cultures of iPS- and ES-derived MPCs were then differentiated for 20 or 27 days in MesenCult™ Osteogenic Differentiation Medium. Strong alkaline phosphatase activity (red stain) and bone mineralization (black stain) were observed at 20 days of osteogenic differentiation, which was further enhanced after 27 days of osteogenic differentiation.

Publications

(4)
Stem cell research {\&} therapy 2020 jan

Scaffold vascularization method using an adipose-derived stem cell (ASC)-seeded scaffold prefabricated with a flow-through pedicle.

T. D\cebski et al.

Abstract

BACKGROUND Vascularization is important for the clinical application of tissue engineered products. Both adipose-derived stem cells (ASCs) and surgical prefabrication can be used to induce angiogenesis in scaffolds. Our aim was to compare the angiogenic potential of ASC-seeded scaffolds combined with scaffold prefabrication with that of non-seeded, non-prefabricated scaffolds. METHODS For prefabrication, functional blood vessels were introduced into the scaffold using a flow-through pedicle system. ASCs were isolated from rat fat deposits. Three-dimensional-printed cylindrical poly-$\epsilon$-caprolactone scaffolds were fabricated by fused deposition modelling. Three groups, each containing six rats, were investigated by using non-seeded, ASC-seeded, and osteogenic induced ASC-seeded scaffolds. In each group, one rat was implanted with two scaffolds in the inguinal region. On the right side, a scaffold was implanted subcutaneously around the inferior epigastric vessels (classic prefabrication group). On the left side, the inferior epigastric vessels were placed inside the prefabricated scaffold in the flow-through pedicle system (flow-through prefabrication group). The vessel density and vascular architecture were examined histopathologically and by $\mu$CT imaging, respectively, at 2 months after implantation. RESULTS The mean vessel densities were 10- and 5-fold higher in the ASC-seeded and osteogenic induced ASC-seeded scaffolds with flow-through prefabrication, respectively, than in the non-seeded classic prefabricated group (p {\textless} 0.001). $\mu$CT imaging revealed functional vessels within the scaffold. CONCLUSION ASC-seeded scaffolds with prefabrication showed significantly improved scaffold vasculogenesis and could be useful for application to tissue engineering products in the clinical settings.
Inflammation 2019 jun

Therapeutic Effects of Mesenchymal Stem Cells Derived From Bone Marrow, Umbilical Cord Blood, and Pluripotent Stem Cells in a Mouse Model of Chemically Induced Inflammatory Bowel Disease.

A. Kagia et al.

Abstract

Acute inflammatory bowel disease (AIBD) is a wide clinical entity including severe gastrointestinal pathologies with common histopathological basis. Epidemiologically increasing diseases, such as necrotizing enterocolitis (NEC), gastrointestinal graft versus host disease (GVHD), and the primary acute phase of chronic inflammatory bowel disease (CIBD), exhibit a high necessity for new therapeutic strategies. Mesenchymal stem cell (MSC) cellular therapy represents a promising option for the treatment of these diseases. In our study, we comparatively assess the efficacy of human MSCs derived from bone marrow (BM), umbilical cord blood (UCB), human embryonic stem cells (ESCs), or human-induced pluripotent stem cells (iPSCs) in a mouse model of chemically induced acute enterocolitis. The laboratory animals were provided ad libitum potable dextrane sulfate sodium solution (DSS) in order to reproduce an AIBD model and then individually exposed intraperitoneally to MSCs derived from BM (BM-MSCs), UCB (UCB-MSCs), ESCs (ESC-MSCs), or iPSCs (iPSC-MSCs). The parameters used to evaluate the cellular treatment efficacy were the animal survival prolongation and the histopathological-macroscopic picture of bowel sections. Although all categories of mesenchymal stem cells led to statistically significant survival prolongation compared to the control group, significant clinical and histopathological improvement was observed only in mice receiving BM-MSCs and UCB-MSCs. Our results demonstrated that the in vivo anti-inflammatory effect of ESC-MSCs and iPSC-MSCs was inferior to that of UCB-MSCs and BM-MSCs. Further investigation will clarify the potential of ESCs and iPSC-derived MSCs in AIBD treatment.
JCI insight 2019 aug

Mesenchymal stromal cells lower platelet activation and assist in platelet formation in vitro.

A. Mendelson et al.

Abstract

The complex process of platelet formation originates with the hematopoietic stem cell, which differentiates through the myeloid lineage, matures, and releases proplatelets into the BM sinusoids. How formed platelets maintain a low basal activation state in the circulation remains unknown. We identify Lepr+ stromal cells lining the BM sinusoids as important contributors to sustaining low platelet activation. Ablation of murine Lepr+ cells led to a decreased number of platelets in the circulation with an increased activation state. We developed a potentially novel culture system for supporting platelet formation in vitro using a unique population of CD51+PDGFRalpha+ perivascular cells, derived from human umbilical cord tissue, which display numerous mesenchymal stem cell (MSC) properties. Megakaryocytes cocultured with MSCs had altered LAT and Rap1b gene expression, yielding platelets that are functional with low basal activation levels, a critical consideration for developing a transfusion product. Identification of a regulatory cell that maintains low baseline platelet activation during thrombopoiesis opens up new avenues for improving blood product production ex vivo.
Cell reports 2019

The Ion Transporter NKCC1 Links Cell Volume to Cell Mass Regulation by Suppressing mTORC1.

W. L. Demian et al.

Abstract

mTORC1 regulates cellular growth and is activated by growth factors and by essential amino acids such as Leu. Leu enters cells via the Leu transporter LAT1-4F2hc (LAT1). Here we show that the Na+/K+/2Cl- cotransporter NKCC1 (SLC12A2), a known regulator of cell volume, is present in complex with LAT1. We further show that NKCC1 depletion or deletion enhances LAT1 activity, as well as activation of Akt and Erk, leading to activation of mTORC1 in cells, colonic organoids, and mouse colon. Moreover, NKCC1 depletion reduces intracellular Na+ concentration and cell volume (size) and mass and stimulates cell proliferation. NKCC1, therefore, suppresses mTORC1 by inhibiting its key activating signaling pathways. Importantly, by linking ion transport and cell volume regulation to mTORC1 function, NKCC1 provides a long-sought link connecting cell volume (size) to cell mass regulation.
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