MesenCult™-ACF软骨细胞分化试剂盒

用于MSCs向软骨细胞分化的无动物成分培养基

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产品号 #（选择产品）

产品号 #05455_C

用于MSCs向软骨细胞分化的无动物成分培养基

产品优势

无动物成分（ACF）配方；
最快需要14天，实现最少3 × 10^5 MSCs高效地软骨分化；
软骨相关转录因子Acan、Col2a、Sox9和Col10a的强表达；肥大相关转录因子Mmp13低表达；
优化的完整无动物成分（ACF）产品线，用于MSC的分离、扩增、冻存和软骨分化

产品组分包括

MesenCult™-ACF软骨分化基础培养基，95 mL
MesenCult™-ACF 20X 软骨分化补充剂， 5 mL

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总览

MesenCult™-ACF软骨分化培养基是一种无动物成分（ACF）的软骨分化培养基，专门用于人原代间充质基质细胞和多能干细胞来源的间充质祖细胞(MSCs；也称为间充质干细胞)分化为软骨生成谱系的细胞，包括软骨细胞。该培养基适用于人骨髓（Bone marrow），脂肪组织（Adipose tissue）和滑膜（Synovium)来源的 MSC 的分化，MSC可在含血清培养基（例如MesenCult™ 扩增试剂盒 (人)[产品号 #05411]）或无动物成分的MesenCult™-ACF Plus Medium培养基[产品号 #05445]）中培养扩增。MesenCult™-ACF软骨分化培养基可诱导最少3 × 10^5个细胞，最早可在第14天实现人MSC高效地软骨分化。

实验数据

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.

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.

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.

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.

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文献 (11)

Endogenous CCN family member WISP1 inhibits trauma-induced heterotopic ossification. G. C.-Y. Hsu et al. JCI insight 2020 jul

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.

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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. Stem cells and development 2020 jul

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.

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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. Biomedicine {\&} pharmacotherapy = Biomedecine {\&} pharmacotherapie 2020 jan

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.

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种属	Human
配方类别	Animal Component-Free, Serum-Free

MesenCult™-ACF软骨细胞分化试剂盒

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产品号 #05455_C

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MesenCult™-ACF软骨细胞分化试剂盒

产品号 #（选择产品）

产品号 #05455_C

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产品说明书及文档

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