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Osteogenic Inducible Small Molecule Compound Kit-1(Powder,With Alizarin Red S and CPC)

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Osteogenic Inducible Small Molecule Compound Kit-1(Powder,With Alizarin Red S and CPC)
Cat.No:
IK-OIN-1
Brand:
Solarbio


货号产品名称规格对应靶点产品描述
ID0170地塞米松10mgGlucocorticoid Receptor地塞米松是成骨诱导的基础试剂,可以激活位于干细胞表面的糖皮质激素受体,显著地增加碱性磷酸酶的活性;可以诱导干细胞向成骨细胞分化。
IA0530维生素C10mgOthers一种重要的营养素和氧化还原剂,在骨盐代谢及骨形成中具有重要作用。在体外,它能增加钙盐的沉积,促进矿化结节的形成,并通过间充质细胞间接调节破骨细胞的分化。维生素C能直接抑制核因子κB受体活化子配体(RANKL)诱导的破骨细胞形成 。
IG1340β-甘油磷酸钠五水合物10mgPhosphatase可以为成骨细胞提供磷酸离子,同时促进生理性钙盐的沉积和钙化,是骨髓基质干细胞产生矿化结节的必要条件。可促进地塞米松对干细胞的诱导分化。
IA5140茜素红S50mg
是一种蒽醌类染料,能与钙盐以鳌和方式形成一种深红色的复合物,用以识别组织细胞的钙盐成分。
IC5411氯化十六烷基吡啶500mg
氯化十六烷基吡啶可用于在茜素红对细胞矿化染色后,使用氯化十六烷基吡啶对矿化结节进行半定量分析。

1. 本套餐精选了3种经典的可用于成骨诱导的基础试剂和高品质的茜素红染料和氯化十六烷基吡啶试剂,均具有良好的生物活性。客户也可以根据自己的实验需求删减或添加其他化合物(如钙离子或者是维生素D。客户可根据自己的需求灵活定制专属Kit

2. 已经过NMRHPLCMS测定,以确保化合物纯度和准确性


基本操作(仅供参考

1.接种骨髓间充质干细胞:取对数生长期的细胞,按照2×104cells/cm2的细胞密度接种至包被后的培养器皿中,于37℃,5% CO2培养环境下培养至融合度60-70%,弃掉上清,加入成骨诱导分化培养基。

2.细胞分化诱导:每2-3天更换成骨诱导培养基,于37℃,5%CO2培养环境下培养,并注意观察细胞形态变化(一般约14~28天左右)。根据细胞钙盐结晶析出和钙质结节形成的情况,决定终止细胞诱导的时间,进行染色鉴定。

3.细胞固定:吸去培养基使用适量1×PBS清洗一次,弃去后取适量4%中性甲醛溶液覆盖培养器皿底面,室温固定30-60 min,弃去固定液再使用1×PBS清洗两次。

4.茜素红染色:加入适量茜素红染液染3~5min,吸去茜素红染液,用1×PBS清洗两次,并加入适量1×PBS避免细胞干燥。

5.诱导评估:显微镜下观察成骨染色效果,并进行图像采集和诱导评估。诱导成功时,钙质结节会与茜素红染料结合后呈现红色或橘红色。

6.半定量分析:显微观察结束后,弃上清,加入氯化十六烷基吡啶溶液,室温下孵育15~60min溶解矿化结节(茜素红),然后在562 nm处检测上清的OD值。


使用本产品的案例(仅供参考

In Vitro

Cell( PDLSCs and GMSCs; 10 nM dexamethasone, 10 mM β-glycerophosphate , and 50 mg/l ascorbic acid; 4 weeks)

Multilineage differentiation assays were applied to detect the pluripotency of PDLSCs and GMSCs.   For osteogenic or adipogenic induction, 2 × 105 cells were seeded in 6-well dishes and cultured in osteogenic medium or adipogenic medium for 4 weeks, then the cells were fixed with 4% paraformaldehyde and stained with Alizarin Red (Solarbio, Beijing, China) or Oil Red O (Solarbio).    ... ...   The osteogenic medium was the complete culture medium supplemented with 10 nM dexamethasone(Solarbio), 10 mM β-glycerophosphate (Solarbio), and 50 mg/l ascorbic acid (Solarbio).  The adipogenic medium was the complete culture medium supplemented with 1 μM dexamethasone (Solarbio), 0.2 mM indomethacin (Solarbio), 0.01 g/l insulin (Solarbio), and 0.5 mM isobutyl-methylxanthine(Solarbio).

来源文献:Jia L, Zhang Y, Ji Y, Li X, Xing Y, Wen Y, Huang H, Xu X. Comparative analysis of lncRNA and mRNA expression profiles between periodontal ligament stem cells and gingival mesenchymal stem cells. Gene. 2019 May 30;699:155-164. doi: 10.1016/j.gene.2019.03.015. Epub 2019 Mar 12. PMID: 30876821.




Remark:
These protocols are for reference only. Solarbio does not independently validate these methods.

Note:
1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.
2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.
3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.
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