Hepatocyte wash buffer

Manufactured by Avantor

Sourced in United States

Hepatocyte wash buffer is a laboratory reagent used to prepare and maintain hepatocytes, which are the primary functional cells of the liver. The buffer is designed to gently wash and remove contaminants from isolated hepatocytes, preserving their viability and functionality for further applications in cell culture and research.

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Lab products found in correlation

Hepatocyte wash buffer, by Thermo Fisher Scientific (2 mentions) Tissue digestion enzymes, by Worthington (2 mentions) Round bottom tube, by Corning (1 mentions)

2 protocols using hepatocyte wash buffer

Tissue Digestion and Cell Isolation Protocol

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Remaining tissue not used for tissue slice analysis (90%) was used for
tissue digestion and cell isolation. Tissue digestion enzymes (Worthington
Biochemical, Lakewood, NJ USA) and conditions are described in the table seen in
Figure 3A. All digestion buffers were
made in 1mL Hepatocyte Wash Buffer (ThermoFisher Scientific) and digested in 5mL
round bottom tubes (Corning). The digestion reaction was neutralized by adding
equal volume (1mL) of 10% fetal bovine serum diluted in Hepatocyte Wash Buffer
(VWR, Radnor, PA USA). Samples were then strained through a 100 uM tube top
filter (Corning) and washed with 500 uL 1X PBS. Cell suspensions were
centrifuged at 1000 RPM (200xG) for 3 minutes and re-suspended in 300 uL MACS
Buffer for cell isolation. This re-suspended sample is referred to as the
“raw digestate”. 20 uL of the raw digestate was removed for
viability and cellular composition analysis. Cells were stained with viability
markers and fluorescent antibodies as described above and reagent specifics can
be found in Supplementary
Table S1. The remaining 280uL of raw digestate was used for cell
isolation.

Vitek R.A., Huang W., Geiger P.G., Heninger E., Lang J.M., Jarrard D.F., Beebe D.J, & Johnson B.P. (2022). Fresh tissue procurement and preparation for multicompartment and multimodal analysis of the prostate tumor microenvironment. The Prostate, 82(7), 836-849.

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Tissue Digestion and Cell Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Remaining tissue not used for tissue slice analysis (90%) was used for tissue digestion and cell isolation. Tissue digestion enzymes (Worthington Biochemical) and conditions are described in the table seen in Figure 3A. All digestion buffers were made in 1 mL Hepatocyte Wash Buffer (Thermo Fisher Scientific) and digested in 5 ml round bottom tubes (Corning). The digestion reaction was neutralized by adding equal volume (1 ml) of 10% fetal bovine serum diluted in Hepatocyte Wash Buffer (VWR). Samples were then strained through a 100 μM tube top filter (Corning) and washed with 500 μl 1X phosphate‐buffered saline (PBS). Cell suspensions were centrifuged at 1000 RPM (200 g) for 3 min and re‐suspended in 300 μl MACS Buffer for cell isolation. This re‐suspended sample is referred to as the “raw digestat.” 20 μl of the raw digestate was removed for viability and cellular composition analysis. Cells were stained with viability markers and fluorescent antibodies as described above and reagent specifics can be found in Table S1. The remaining 280 μl of raw digestate was used for cell isolation.

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