Tuesday, June 28, 2016

Cells Detachment

Easy, Gentle and Complete

I often get asked if trypsin is the best reagent to detach primary and stem cells. The problem with a trypsin is that it could lyse a percentage of the cells.

We recommend our DetachinTM. It offers the following benefits:
  • Gentle and rapid detachment.
  • Maximum cell viability.
  • Effective on a wide range of cells.
  • No mammalian or bacterial byproducts.
  • Stable at 4°C for 2 months.
  • No need to wash detached cells.
Here's a recent publication referencing the use of Detachin: Yu Hou, Qi Feng, Miao Xu, Guo-sheng Li, Xue-na Liu, Zi Sheng, Hai Zhou, Ji Ma, Yu Wei,1 Yuan-xin Sun,Ying-yi Yu, Ji-hua Qiu, Lin-lin Shao, Xin-guang Liu, Ming Hou, and Jun Peng. High-dose dexamethasone corrects impaired myeloid-derived suppressor cell function via Ets1 in immune thrombocytopenia. Blood 2016 :blood-2015-10-674531; doi:10.1182/blood-2015-10-674531 ...Both PBMCs and splenocytes were cultured in complete medium for 7 days. Each culture was supplemented with recombinant human IL-6 (10 ng/mL; R&D Systems, Minneapolis, MN) and granulocyte macrophage-colony-stimulating factor (10 ng/mL; R&D Systems) in the presence or absence of DXM and incubatedin humidified air with 5%CO2 at 37°C. Cells cultured in medium alone were runin parallel as controls. Adherent cells were removed using a nonprotease cell detachment solution Detachin (Neuromics, Edina, MN). CD331 cells were isolated by anti-CD33 magnetic microbeads and LS column separation (Miltenyi Biotec, Bergisch Gladbach, Germany), per manufacturer's instructions. The purity of the isolated cells was found to be higher than 90% by flow cytometry

Figure 1. The number of MDSCs and their expression of Arg-1/iNOS in the peripheral blood. (A) The representative scatter-gram of CD11b1CD331HLA-DRlow cells within the gate of PBMCs. Histograms of Arg-1 and iNOS in CD11b1CD331HLA-DRlow cells from healthy control patients and patients with ITP before any treatment was initiated. (B) The percentage of CD11b1CD331HLA-DRlow cells in hemolyzed whole blood from primary active patients with ITP (n 5 21) and healthy control patients (n 5 18). (C-D) The expression (mean fluorescence intensity) of Arg-1 (C) and iNOS (D) in circulating MDSCs compared between patients with ITP (n 5 21) and control patients (n 5 18). Significance between the 2 groups was determined by Student-Newman-Keuls test.
We are working hard to find new ways for you to optimize your cell based assays.

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