MesenCult™-ACF Chondrogenic Differentiation Kit

Animal component-free medium for the differentiation of MSCs into chondrocytes

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MesenCult™-ACF Chondrogenic Differentiation Kit

Animal component-free medium for the differentiation of MSCs into chondrocytes

100 mL
Catalog #05455
249 USD

Required Products

Overview

MesenCult™-ACF Chondrogenic Differentiation Medium is animal component-free (ACF) and specifically formulated for the in vitro differentiation of human mesenchymal stromal cells (MSCs; also known as mesencyhymal stem cells) into chondrogenic lineage cells, including chondrocytes. This medium is suitable for the differentiation of human bone marrow (BM)-, adipose tissue (AT)- and synovium (S)-derived MSCs previously culture-expanded in serum-containing medium (e.g. MesenCult™ Proliferation Kit [Catalog #05411]) or animal component-free MesenCult™-ACF Plus Medium [Catalog #05445]). MesenCult™-ACF Chondrogenic Differentiation Medium induces robust chondrogenic differentiation of human MSCs with as few as 3 x 10^5 cells and as early as day 14.
Advantages:
• Animal component-free (ACF) formulation
• Robust chondrogenic differentiation with as few as 3 x 10^5 MSCs and as early as day 14
• Strong expression of chondrogenic transcripts - Acan, Col2a, Sox9 and Col10a; low expression of hypertrophic transcript Mmp13
• Completes optimized ACF workflow for MSC isolation, expansion, cryopreservation and chondrogenic differentiation
Components:
  • MesenCult™-ACF Chondrogenic Differentiation Basal Medium, 95 mL
  • MesenCult™-ACF 20X Chondrogenic Differentiation Supplement, 5 mL
Subtype:
Specialized Media
Cell Type:
Chondrocytes; Mesenchymal Stem and Progenitor Cells
Species:
Human
Application:
Cell Culture; Differentiation
Brand:
MesenCult
Area of Interest:
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

MesenCult™-ACF Chondrogenic Differentiation Medium Induces Robust Chondrogenic Differentiation of Human MSCs

Figure 1. MesenCult™-ACF Chondrogenic Differentiation Medium Induces Robust Chondrogenic Differentiation of Human MSCs

Human BM-derived MSCs were cultured in MesenCult™-ACF Medium then differentiated to the chondrogenic lineage using MesenCult™-ACF Chondrogenic Differentiation Medium. Robust chondrogenic differentiation was observed (A) starting with as few as 3 x 105 MSCs, or (B) when differentiating for just 14 days starting with 5 x 105 MSCs.

Chondrogenic Differentiation of Human MSCs Is More Robust With Fewer Hypertrophic Chondrocytes Using MesenCult™-ACF Chondrogenic Differentiation Medium Compared to Competitor Media

Figure 2. Chondrogenic Differentiation of Human MSCs Is More Robust With Fewer Hypertrophic Chondrocytes Using MesenCult™-ACF Chondrogenic Differentiation Medium Compared to Competitor Media

Human BM-derived MSCs culture-expanded for up to two passages in MesenCult™-ACF Medium, serum-based medium, or one of two commercially available media (Competitor 1 (Ex1) and Competitor 2 (Ex2)), were then differentiated to the chondrogenic lineage starting with 5 x 105 MSCs and either using MesenCult™-ACF Chondrogenic Differentiation Medium or one of several commercially available chondrogenic differentiation media (Ch1, Ch2 or Ch3) for 21 days. More robust and uniform chondrogenic differentiation was observed when the MSCs were differentiated in MesenCult™-ACF Chondrogenic Differentiation Medium compared to the other commercially available chondrogenic differentiation media (Ch1, Ch2 and Ch3), irrespective of the expansion medium used to culture the MSCs prior to differentiation. Cultures differentiated using MesenCult™-ACF Chondrogenic Differentiation Medium displayed an abundance of isogenous groups (yellow arrows), suggesting there is proliferation of differentiating chondrocyte progenitors. Few hypertrophic chondrocytes (black arrows) are seen in cultures differentiated with MesenCult™-ACF Chondrogenic Differentiation Medium, suggesting the maintenance of chondrogenic activity throughout the culturing period.

MesenCult™-ACF Chondrogenic Differentiation Medium Induces Stronger and More Sustained Chondrogenic Transcript Expression Compared to Competitor Media

Figure 3. MesenCult™-ACF Chondrogenic Differentiation Medium Induces Stronger and More Sustained Chondrogenic Transcript Expression Compared to Competitor Media

Human BM-derived MSCs expanded in (A) MesenCult™-ACF Medium, (B) a serum-based medium or (C) Competitor 2 (Ex2) medium, were differentiated for 21 days with MesenCult™-ACF Chondrogenic Differentiation Medium and Competitor 2 (Ch2) chondrogenic differentiation medium. Regardless of the expansion medium initially used to culture the MSCs, differentiation using MesenCult™-ACF Chondrogenic Differentiation Medium led to a substantial up-regulation of the chondrogenic transcripts compared to Ch2. In addition, expression of the terminally-differentiated hypertrophic transcript Mmp13 was higher for Ch2 differentiated cultures compared to cultures differentiated with MesenCult™-ACF Chondrogenic Differentiation Medium.

Mouse MSCs Cultured in MesenCult™-ACF Chondrogenic Differentiation Medium Differentiate to Chondrocytes

Figure 4. Mouse MSCs Cultured in MesenCult™-ACF Chondrogenic Differentiation Medium Differentiate to Chondrocytes

Mouse compact bone-derived MSCs were cultured using the MesenCult™ Proliferation Kit with MesenPure™ (Mouse, Catalog #05512) for 2 passages then differentiated by pellet culture with MesenCult™-ACF Chondrogenic Differentiation Medium for 21 days under normoxic (20% O2) conditions. Strong chondrogenic differentiation is indicated by dark-blue staining of the cartilage extracellular matrix and an abundance of isogenous chondrocyte groups.

Human MSC Chondrogenic Differentiation With AggreWell™800 Plates

Figure 5. Human MSC Chondrogenic Differentiation With AggreWell™800 Plates

Using centrifugation, 1 x 10^6 human MSCs were distributed evenly among 800 µm microwells in one well of an AggreWell™800 plate. Small aggregates of only ~3,300 cells per pellet were then differentiated to chondrocytes using MesenCult™-ACF Chondrogenic Differentiation Medium for 21 days under normoxic (20% O2) conditions.

Procedure Overview: Human MSC Chondrogenic Differentiation Using MesenCult™-ACF Chondrogenic Differentiation Medium

Figure 6. Procedure Overview: Human MSC Chondrogenic Differentiation Using MesenCult™-ACF Chondrogenic Differentiation Medium

Aggregate culture is a useful method for inducing chondrogenic differentiation of human BM- and adipose-derived MSCs in a three-dimensional in vitro culture environment. MSCs are efficiently differentiated to the chondrogenic lineage using MesenCult™-ACF Chondrogenic Differentiation Medium in 14 - 21 days with 3 - 5 x 105 cells.

Publications

(11)
Stem cells and development 2020 jul

Human Umbilical Cord Mesenchymal Stem Cells Attenuate Abdominal Aortic Aneurysm Progression in Sprague-Dawley Rats: Implication of Vascular Smooth Muscle Cell Phenotypic Modulation.

H. Wen et al.

Abstract

Abdominal aortic aneurysm (AAA) is life-threatening, for which efficient nonsurgical treatment strategy has not been available so far. Several previous studies investigating the therapeutic effect of mesenchymal stem cells (MSCs) in AAA indicated that MSCs could inhibit aneurysmal inflammatory responses and extracellular matrix destruction, and suppress aneurysm occurrence and expansion. Vascular smooth muscle cell (VSMC) phenotypic plasticity is reported to be predisposed in AAA initiation and progression. However, little is known about the effect of MSCs on VSMC phenotypic modulation in AAA. In this study, we investigate the therapeutic efficacy of umbilical cord mesenchymal stem cells (UC-MSCs) in elastase-induced AAA model and evaluate the effect of UC-MSC on VSMC phenotypic regulation. We demonstrate that the intravenous injection of UC-MSC attenuates elastase-induced aneurysmal expansion, reduces elastin degradation and fragmentation, inhibits MMPs and TNF-$\alpha$ expression, and preserves and/or restores VSMC contractile phenotype in AAA. Taken together, these results highlight the therapeutic and VSMC phenotypic modulation effects of UC-MSC in AAA progression, which further indicates the potential of applying UC-MSC as an alternative treatment candidate for AAA.
JCI insight 2020 jul

Endogenous CCN family member WISP1 inhibits trauma-induced heterotopic ossification.

G. C.-Y. Hsu et al.

Abstract

Heterotopic ossification (HO) is defined as abnormal differentiation of local stromal cells of mesenchymal origin, resulting in pathologic cartilage and bone matrix deposition. Cyr61, CTGF, Nov (CCN) family members are matricellular proteins that have diverse regulatory functions on cell proliferation and differentiation, including the regulation of chondrogenesis. However, little is known regarding CCN family member expression or function in HO. Here, a combination of bulk and single-cell RNA sequencing defined the dynamic temporospatial pattern of CCN family member induction within a mouse model of trauma-induced HO. Among CCN family proteins, Wisp1 (also known as Ccn4) was most upregulated during the evolution of HO, and Wisp1 expression corresponded with chondrogenic gene profile. Immunohistochemistry confirmed WISP1 expression across traumatic and genetic HO mouse models as well as in human HO samples. Transgenic Wisp1LacZ/LacZ knockin animals showed an increase in endochondral ossification in HO after trauma. Finally, the transcriptome of Wisp1-null tenocytes revealed enrichment in signaling pathways, such as the STAT3 and PCP signaling pathways, that may explain increased HO in the context of Wisp1 deficiency. In sum, CCN family members, and in particular Wisp1, are spatiotemporally associated with and negatively regulate trauma-induced HO formation.
Biomedicine {\&} pharmacotherapy = Biomedecine {\&} pharmacotherapie 2020 jan

Bone marrow mesenchymal stem cells alleviate the daunorubicin-induced subacute myocardial injury in rats through inhibiting infiltration of T lymphocytes and antigen-presenting cells.

Q. Chen et al.

Abstract

INTRODUCTION Bone marrow mesenchymal stem cells (BMSCs) have been extensively investigated from a perspective on cardiac regeneration therapy. The current study aimed to investigate the protective effect conferred by BMSCs in subacute myocardial injury, and to identify an appropriate BMSC reinfusion time. METHODS BMSCs were isolated from human bone marrow blood. Daunorubicin (DNR)-induced subacute myocardial models were subsequently established. The rats with DNR-induced subacute myocardial injury were injected with dexrazoxane (DZR) and/or BMSCs at varying time points, after which cardiac function was evaluated by assessing left ventricular ejection fraction (LVEF) and fraction shortening (FS). The myocardial structural changes were analyzed, after which the levels of CD3 and human leukocyte antigen DR (HLA-DR) were examined to further validate the mechanism by which BMSCs could influence subacute myocardial injury. RESULTS BMSCs combined with DZR treatment enhanced the cardiac function of rats with DNR-induced myocardial injury, as reflected by increased LVEF and FS. DNR-induced myocardial injuries were mitigated via the application of BMSCs combined with treatment of DZR, accompanied by diminished infiltration or vacuolization. Moreover, BMSCs were observed to alleviate infiltration of T lymphocyte and antigen-presenting cells, as evidenced by reduced expression of CD3 and HLA-DR. CONCLUSION Taken together, this study demonstrates that BMSCs could protect against DNR-induced myocardial injury, especially in the first three days of DNR administration. BMSCs combined with DZR exert a better therapeutic effect, but there are individual differences.
Scientific reports 2020 feb

Thalidomide Inhibits Human iPSC Mesendoderm Differentiation by Modulating CRBN-dependent Degradation of SALL4.

D. G. Belair et al.

Abstract

Exposure to thalidomide during a critical window of development results in limb defects in humans and non-human primates while mice and rats are refractory to these effects. Thalidomide-induced teratogenicity is dependent on its binding to cereblon (CRBN), the substrate receptor of the Cul4A-DDB1-CRBN-RBX1 E3 ubiquitin ligase complex. Thalidomide binding to CRBN elicits subsequent ubiquitination and proteasomal degradation of CRBN neosubstrates including SALL4, a transcription factor of which polymorphisms phenocopy thalidomide-induced limb defects in humans. Herein, thalidomide-induced degradation of SALL4 was examined in human induced pluripotent stem cells (hiPSCs) that were differentiated either to lateral plate mesoderm (LPM)-like cells, the developmental ontology of the limb bud, or definitive endoderm. Thalidomide and its immunomodulatory drug (IMiD) analogs, lenalidomide, and pomalidomide, dose-dependently inhibited hiPSC mesendoderm differentiation. Thalidomide- and IMiD-induced SALL4 degradation can be abrogated by CRBN V388I mutation or SALL4 G416A mutation in hiPSCs. Genetically modified hiPSCs expressing CRBN E377V/V388I mutant or SALL4 G416A mutant were insensitive to the inhibitory effects of thalidomide, lenalidomide, and pomalidomide on LPM differentiation while retaining sensitivity to another known limb teratogen, all-trans retinoic acid (atRA). Finally, disruption of LPM differentiation by atRA or thalidomide perturbed subsequent chondrogenic differentiation in vitro. The data here show that thalidomide, lenalidomide, and pomalidomide affect stem cell mesendoderm differentiation through CRBN-mediated degradation of SALL4 and highlight the utility of the LPM differentiation model for studying the teratogenicity of new CRBN modulating agents.
Bioactive materials 2020 dec

An all-silk-derived functional nanosphere matrix for sequential biomolecule delivery and in situ osteochondral regeneration.

W. Zhang et al.

Abstract

Endogenous repair of osteochondral defect is usually limited by the insufficient number of cells in the early stage and incomplete cell differentiation in the later stage. The development of drug delivery systems for sequential release of pro-migratory and pro-chondrogenic molecules to induce endogenous bone marrow-derived mesenchymal stem cells (BMSCs) recruitment and chondrogenic differentiation is highly desirable for in situ osteochondral regeneration. In this study, a novel, all-silk-derived sequential delivery system was fabricated by incorporating the tunable drug-loaded silk fibroin (SF) nanospheres into a SF porous matrix. The loading efficiency and release kinetics of biomolecules depended on the initial SF/polyvinyl alcohol (PVA) concentrations (0.2{\%}, 1{\%} and 5{\%}) of the nanospheres, as well as the hydrophobicity of the loaded molecules, resulting in controllable and programmed delivery profiles. Our findings indicated that the 5{\%} nanosphere-incorporated matrix showed a rapid release of E7 peptide during the first 120 h, whereas the 0.2{\%} nanosphere-incorporated matrix provided a slow and sustained release of Kartogenin (KGN) longer than 30 days. During in vitro culture of BMSCs, this functional SF matrix incorporated with E7/KGN nanospheres showed good biocompatibility, as well as enhanced BMSCs migration and chondrogenic differentiation through the release of E7 and KGN. Furthermore, when implanted into rabbit osteochondral defect, the SF nanosphere matrix with sequential E7/KGN release promoted the regeneration of both cartilage and subchondral bone. This work not only provided a novel all-silk-derived drug delivery system for sequential release of molecules, but also a functional tissue-engineered scaffold for osteochondral regeneration.
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