Effect of purified fractions from cell culture supernate of high‐density pre‐B acute lymphoblastic leukemia cells (ALL3) on the growth of ALL3 cells at low density

• Published in: Electrophoresis Vol. 38; no. 3-4; pp. 417 - 428
• Main Authors: Patel, Sapan J., Darie, Costel C., Clarkson, Bayard D.
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 01-02-2017
• Subjects: Aberration; Acute lymphoblastic leukemia; Biochemistry; Bioinformatics; Biotechnology; Cell Communication; Cell Culture Techniques; Cell Line, Tumor; Cell Proliferation; Cells, Cultured - drug effects; Collective Behavior; Culture Media, Conditioned - metabolism; Culture Media, Conditioned - pharmacology; Density; Humans; Leukemia; Leukemias; Mass spectrometry; Nanostructure; Precursor Cell Lymphoblastic Leukemia-Lymphoma - metabolism; Protein purification; Proteins; Proteins - analysis; Proteins - metabolism; Proteins - secretion; Proteome - analysis; Proteome - drug effects; Proteome - metabolism; Proteome - secretion; Purification; Quorum Sensing; Secretome; Serum Removal; Secretome; Quorum Sensing; Serum Removal; Acute lymphoblastic leukemia; Collective Behavior; Protein purification
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/elps.201600399

The mechanisms underlying the aberrant growth and interactions between cells are not understood very well. The pre‐B acute lymphoblastic leukemia cells directly obtained from an adult patient grow very poorly or do not grow at all at low density (LD), but grow better at high starting cell density (HD). We found that the LD ALL3 cells can be stimulated to grow in the presence of diffusible, soluble factors secreted by ALL3 cells themselves growing at high starting cell density. We then developed a biochemical purification procedure that allowed us to purify the factor(s) with stimulatory activity and analyzed them by nanoliquid chromatography‐tandem mass spectrometry (nanoLC‐MS/MS). Using nanoLC‐MS/MS we have identified several proteins which were further processed using various bioinformatics tools. This resulted in eight protein candidates which might be responsible for the growth activity on non‐growing LD ALL3 cells and their involvement in the stimulatory activity are discussed.