B-27 supplement (50X)，minus vitamin A

Product Description

B-27 supplement (50X)，minus vitamin A is an optimized serum-free additive based on the original B-27 with vitamin A (retinyl acetate) removed. Vitamin A can be converted to retinoic acid, which induces the differentiation of neural stem cells. Vitamin A-free B-27 in combination with a suitable basal medium is suitable for the expansion of pluripotent stem cell (PSC)-derived neural progenitor cells, as well as for the growth and maintenance of the activity of primary neural stem and neural progenitor cells, which can be cultured in suspension without inducing their differentiation.

Production process: This product is prepared by using Water-For-Injection (WFI), and the production procedure is strictly in accordance with the cGMP management regulations, and is completed in the clean room for the record of in vitro diagnostic reagents. The company has obtained ISO9001 quality system certification.

Product Advantage

• Removes vitamin A to support neural stem cell expansion as well as specific differentiation processes;
• Increasing the rate of neural stem cell proliferation and viability;
• Maintenance of neural stem cell phenotype and differentiation characteristics;
• Serum-free culture system with optimized formulation;
• Prepared with water for injection, produced in compliance with cGMP regulations, stable process, small batch variation；
• Raw materials are all self-produced or strictly tested and purified low endotoxin high purity pharmaceutical grade raw materials;
• Strict quality testing procedures, much higher than the required indicators in the same industry, the product performance is outstanding, the indicators are better than the international;
• Imported material aseptic packaging bottles, in line with cGMP management regulations, excellent airtightness and internal coating protection, greatly ensuring the stability of the product.

Product packaging form: PWL043-1 is 10ML per bottle; PWL043-2 is 100ML per bottle.

Performance

1. Detection of the effect of promoting mouse neural stem cell proliferation

Methods: Meiluncell Neural-27 (w/o vitamin A) and company G B-27 (w/o vitamin A) were used to culture neural stem cells (NSC) extracted from the fetal brains of C57/BL-6 mice. Cell counting and survival rate detection were performed 72 hours later.

Results: Neurospheres cultured with Meiluncell Neural-27 (w/o vitamin A) proliferated more cells and had a survival rate similar to the control. And the growth status of NSC is good.

Figure 1: Neural-27 supplement (50X), removal of vitamin A to culture NSCs, comparison of cell number and survival rate

Figure 2: Neural-27 supplement (50X), removal of vitamin A to culture NSCs, comparison of cell growth status 

2. Detection of the effect of promoting C17.2 cell proliferation

Methods: Meiluncell Neural-27 (vitamin A removed) and company G B-27 (w/o vitamin A) were used to culture C17.2 cells respectively, and cell counting and survival rate were detected after 72 hours.

Results: C17.2 cells cultured with Meiluncell Neural-27 (w/o vitamin A) proliferated more cells and had a survival rate similar to the control. And the C17.2 cells were in good growth status.

Figure 3: Neural-27 supplement (50X), removal of vitamin A to culture C17.2, comparison of cell number and survival rate

Figure 4: Neural-27 supplement (50X), removal of vitamin A to culture C17.2, comparison of cell growth state

3. Purity and differentiation detection of mouse neural stem cells after culture

Methods: Meiluncell Neural-27 (w/o vitamin A) was used to culture NSC cells, and after a period of time, immunohistochemical identification of purity and induced differentiation was performed.

Results: NSC cells cultured using Meiluncell Neural-27 (w/o vitamin A) were labeled with Nestin on their surface >75%, and the immunohistochemistry results were positive. It shows that Meiluncell Neural-27 (w/o vitamin A) can maintain the expression of neural stem cell surface markers.

After induced differentiation of NSC cells cultured with Meiluncell Neural-27 (w/o vitamin A), the surface labeling of β-Tubulin (neuron-specific protein) was >10%, and the surface labeling of GFAP (astrocyte-specific protein) was >50%, and the immunohistochemistry results were positive. It shows that Meiluncell Neural-27 (w/o vitamin A) can maintain the differentiation characteristics of neural stem cells.

Figure 5: Immunohistochemical identification results of NSC purity

Figure 6: Immunohistochemical identification results of NSC induced differentiation

Shipping and Storage 

• Storage：Store at -20°C away from light for 12 months
• Shipment：< 0°C

Usage Statement

Research Use Only (RUO)