Hexadimethrine bromide (Polybrene) is a cationic polymer used routinely to enhance the efficiency of retrovirus vector-mediated gene transfer^[1][2].

Hexadimethrine bromide inhibits human mesenchymal stem cell proliferation during lentiviral transduction. Hexadimethrine bromide is considered non-toxic at low concentrations, but has been found to negatively affect cell proliferation in some cell types at concentrations greater than 10 µg/mL. Trypsinized cells exposed to Hexadimethrine bromide are visibly larger in size when viewed under the microscope^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

C₁₅H₃₆Br₂N₂

28728-55-4

海(地)美溴铵

Room temperature in continental US; may vary elsewhere.

4°C, stored under nitrogen, away from moisture

*In solvent : -80°C, 6 months; -20°C, 1 month (stored under nitrogen, away from moisture)

H₂O : ≥ 125 mg/mL

DMSO : 5 mg/mL (Need ultrasonic and warming)

* “≥” means soluble, but saturation unknown.

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： PBS

  Solubility: 100 mg/mL (Infinity mM); Clear solution; Need ultrasonic
• 2.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 0.5 mg/mL (Infinity mM); Clear solution

  此方案可获得 ≥ 0.5 mg/mL (Infinity mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 5.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 3.

  请依序添加每种溶剂： 10% DMSO    90% (20% SBE-β-CD in saline)

  Solubility: ≥ 0.5 mg/mL (Infinity mM); Clear solution

  此方案可获得 ≥ 0.5 mg/mL (Infinity mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 5.0 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 4.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: 0.5 mg/mL (Infinity mM); Clear solution; Need warming

  此方案可获得 0.5 mg/mL (Infinity mM) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 5.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. Lin P, et al. Polybrene inhibits human mesenchymal stem cell proliferation during lentiviral transduction. PLoS One. 2011;6(8):e23891.

  [2]. Abe A, et al. Polybrene increases the efficiency of gene transfer by lipofection. Gene Ther. 1998;5(5):708-711.

Trypsinized cells are washed and resuspended in medium at 2×10⁴ cells/mL with or without rhFGF-2 (final concentration=10 ng/mL, PeproTech) and with or without Hexadimethrine bromide at a final Hexadimethrine bromide concentration of 1, 4, or 8 µg/mL. The different conditions are seeded in 96-well plates at 1×10³ cells in 50 µL per well in triplicate and cultured at 37°C, 5% CO₂. After 6, 9, or 24 hr, the medium is changed and subsequent medium changes occurred every 3-4 days with 50 µL of ± FGF-2 medium (10 ng/mL). Plates are harvested on day 4, 7, 14, and 21 by removing the medium and placing the plates in the -80°C freezer until the day of analysis. The CyQUANT assay is then performed on the wells^[1].

上海金畔生物科技有限公司 has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Lin P, et al. Polybrene inhibits human mesenchymal stem cell proliferation during lentiviral transduction. PLoS One. 2011;6(8):e23891.

  [2]. Abe A, et al. Polybrene increases the efficiency of gene transfer by lipofection. Gene Ther. 1998;5(5):708-711.