The biological buffers used in cell therapy

 

During cell therapy, biological buffers are sometimes used to separate cells for culture. The following are the biological buffers used in cell therapy:

Further Reading: The application of Cell Therapy

TRIS-HCl:

  • For Parkinson's Disease, used to detection of Tyrosine Hydroxylase in Differentiated Cells by Western Analysis: The membrane was then blocked for at least 2 hours at room temperature with gentle agitation in 10% BLOTTO (10% nonfat milk powder in TBST [50 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.1% Triton X-100]). [1]
  • Used in estern blot analysis : Brain tissues and co-cultured cells were dissolved in ice-cold lysis buffer (20 mM Tris–HCl, pH 7.5, 1 mM EDTA, 5 mM MgCl2, 1 mM dithiothretol, 0.1 mM phenylmethylsulfonyl fluoride plus protease inhibitor cocktail. The lysates were centrifuged.[2]
  • To preparation of fluorescently labeled FCS: The sample was then added to 0.5 g of FITC dissolved in 5 ml DMSO. After vigorous shaking for 10 min, the reaction was incubated at 4°C for 16 h and then stopped by addition of 0.1 volumes of 1 M Tris–HCl buffer (pH 8.0).[3]
  • For Tumor Immunotherapy: Manganese chloride (MnCl2) solution was firstly added into OVA solution and incubated for 20 minutes under stirring. DAP solution was dropped into the above solution and the pH of mixture was adjusted to pH 7.8-8 by tris (hydroxymethyl) aminomethane-hydrogen chloride (Tris-HCl) buffer.[4]
  • Used in the treat hepatocarcinoma cells: Enzyme assay for artificial substrate For the MMP-1 (collagenase-1) assay, enzyme activities were assayed in a reaction buffer [300 mM NaCl, 10 mM CaCl2, 0.005% Brij35, 0.01% NaN3, and 50 mM Tris-HCl (pH 7.5)] using 20 μM MOAc-Pro-Leu-Gly-Leu-A2pr(Dnp)-Ala-Arg-NH2 as the substrate (90 min incubation at 25°C).[5]
  • Application for Parkinsonism, the PVDF membranes were washed with buffer (0.05% Triton and 0.9% NaCl in 50 mM Tris-HCl, pH 7.3), immersed in the blocking solution for 60 minutes, and then reacted with secondary antibodies at room temperature for 1 hour.[6]
  • Used for Antigen-Targeted Cancer Immunotherapy, Preparation of Recombinant Large HSPs and Tumor Protein Antigen, in Lysis buffer, Binding buffer, Wash buffer and Elution buffer.[7]
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TRIS:

  • Used to detect the role of exosomes derived from bone marrow mesenchymal stem cells (BM-MSCs) and NPCs in their interaction with corresponding cells. BM-MSCs, NPCs and exosomes derived from the two kinds of cells were lysed using RIPA (Beyotime Biotechnology) containing 50 mM Tris (pH 7.4), 1% Triton X-100, 150 mM NaCl, 1% sodium deoxycholate, 0.1% SDS, EDTA, sodium orthovanadate, sodium fluoride, leupeptin and 1 mM PMSF.[8]
  • Assessing Stem Cell DNA Integrity for Cardiac Cell Therapy, for the cell lysis buffer, dissolve 100 mL of lysis buffer (2.5 M NaCl, 0.1 M EDTA, 10 mM Tris, 1% Triton X-100) in deionized water (DIH2O). Then, add 10 M NaOH to adjust the pH to 10.0. Cool the buffer to 4 °C.[9]
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CHAPS:

  • Used in Isolation of HLA ligands. HLA class I molecules were isolated using standard immunoafnity purifcation. Snap-frozen GSC pellets or snap-frozen primary glioblastoma tissue samples were lysed in 10 mM CHAPS/PBS containing 1× protease inhibitor.[10]
  • For a potent apo-IDO1 inhibitor cancer immunotherapy. Recombinant IDO1 Activity Assay Recombinant IDO1 activity at 25°C or 37°C was measured as previously described: 100 nM holo-IDO1 in 100 mM potassium phosphate buffer (pH 7.2) plus 1 mM CHAPS supplemented with 0.5% (v:v) catalase was added (25 µl) to inhibitors in DMSO carrier (0.5 µl DMSO volume) and allowed to incubate for the indicated time and temperature (varied).[11]
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Reference:

[1]https://stemcellsjournals.onlinelibrary.wiley.com/doi/full/10.1634/stemcells.2008-0074

[2]https://doi.org/10.1111/j.1471-4159.2008.05589.x

[3]https://www.sciencedirect.com/science/article/pii/S1525001604000772

[4]https://pubs.acs.org/doi/abs/10.1021/acsnano.9b05974

[5]https://faseb.onlinelibrary.wiley.com/doi/full/10.1096/fj.04-2126com

[6]https://stemcellsjournals.onlinelibrary.wiley.com/doi/full/10.1634/stemcells.2005-0053

[7]https://link.springer.com/protocol/10.1007%2F978-1-4939-7477-1_25

[8]https://link.springer.com/article/10.1186/s13287-017-0563-9

[9]https://www.jove.com/t/58971/assessing-stem-cell-dna-integrity-for-cardiac-cell-therapy

[10]https://link.springer.com/article/10.1007/s00401-018-1836-9

[11]https://www.authorea.com/doi/full/10.22541/au.159863181.10139431

發佈日期:2021.09.24