Disaggregation of human solid tumours by combined mechanical and enzymatic methods

TY - JOUR

T1 - Disaggregation of human solid tumours by combined mechanical and enzymatic methods

AU - Engelholm, S A

AU - Spang-Thomsen, M

AU - Brünner, N

AU - Nøhr, I

AU - Vindeløv, L L

N1 - Keywords: Animals; Cell Separation; DNA, Neoplasm; Flow Cytometry; Humans; Mice; Mice, Nude; Neoplasm Transplantation; Neoplasms, Experimental; Time Factors; Trypsin; Tumor Stem Cell Assay

PY - 1985

Y1 - 1985

N2 - Two combined mechanical and enzymatic disaggregation techniques and a simple mechanical disaggregation procedure were compared. The combined procedures involved a mechanical comminution of the tumour tissue followed by incubation in trypsin. In one method, the tissue was subjected to long-term trypsinization at 4 degrees C, and in the other procedure, repeated short-term trypsinization at 37 degrees C was applied. The results were compared in terms of the yield of viable cells, plating efficiency, the ability to produce tumours in nude mice, and DNA distribution as measured by flow cytometry. The combined techniques provided reproducible cell yields of 2-10 X 10(7) viable cells g-1 of tissue, whereas only a small number of tumour cells was produced by the mechanical method. DNA analysis demonstrated that only the long-term trypsinization procedure resulted in a representative cell yield from all the tumours tested.

AB - Two combined mechanical and enzymatic disaggregation techniques and a simple mechanical disaggregation procedure were compared. The combined procedures involved a mechanical comminution of the tumour tissue followed by incubation in trypsin. In one method, the tissue was subjected to long-term trypsinization at 4 degrees C, and in the other procedure, repeated short-term trypsinization at 37 degrees C was applied. The results were compared in terms of the yield of viable cells, plating efficiency, the ability to produce tumours in nude mice, and DNA distribution as measured by flow cytometry. The combined techniques provided reproducible cell yields of 2-10 X 10(7) viable cells g-1 of tissue, whereas only a small number of tumour cells was produced by the mechanical method. DNA analysis demonstrated that only the long-term trypsinization procedure resulted in a representative cell yield from all the tumours tested.

M3 - Journal article

C2 - 3966975

SN - 0007-0920

VL - 51

SP - 93

EP - 98

JO - The British journal of cancer. Supplement

JF - The British journal of cancer. Supplement

IS - 1

ER -