False leukemia–lymphoma cell lines: an update on over 500 cell lines

Human leukemia–lymphoma (LL) cell lines represent an extremely important resource for research in a variety of fields and disciplines. As the cell lines are used as in vitro model systems in lieu of primary cell material, it is crucial that the cells in the culture flasks faithfully correspond to the purported objects of study. Obviously, proper authentication of cell line derivation and precise characterization are indispensable requirements to use as model systems. A number of studies has shown an unacceptable level of LL cell lines to be false. We present here the results of authenticating a comprehensively large sample (n = 550) of LL cell lines mainly by DNA fingerprinting and cytogenetic evaluation. Surprisingly, near-identical incidences (ca 15%) of false cell lines were observed among cell lines obtained directly from original investigators (59/395: 14.9%) and from secondary sources (23/155: 14.8%) implying that most cross-contamination is perpetrated by originators, presumably during establishment. By comparing our data with those published, we were further able to subclassify the false cell lines as (1) virtual: cross-contaminated with and unretrievably overgrown by other cell lines during initiation, never enjoying independent existence; (2) misidentified: cross-contaminated subsequent to establishment so that an original prototype may still exist; or (3) misclassified: unwittingly established from an unintended (often normal) cell type. Prolific classic leukemia cell lines were found to account for the majority of cross-contaminations, eg CCRF-CEM, HL-60, JURKAT, K-562 and U-937. We discuss the impact of cross-contaminations on scientific research, the reluctance of scientists to address the problem, and consider possible solutions. These findings provide a rationale for mandating the procurement of reputably sourced LL cell lines and their regular authentication thereafter.

[1]  C. Alvarado,et al.  Two new acute lymphoblastic leukemia cell lines with early B-cell phenotypes , 1982 .

[2]  H. Oda,et al.  Establishment of a human megakaryoblastic cell line (T-33) from chronic myelogenous leukemia in megakaryoblastic crisis. , 1988, Cancer research.

[3]  K. Kita,et al.  A novel Ph1 chromosome positive cell line established from a patient with chronic myelogenous leukemia in blastic crisis. , 1985, Leukemia research.

[4]  M. Caligiuri,et al.  Comprehensive cytogenetic and molecular genetic characterization of the TI-1 acute myeloid leukemia cell line reveals cross-contamination with K-562 cell line. , 2002, Blood.

[5]  Ann C. Weller,et al.  Betrayers of the Truth , 1984 .

[6]  V. Diehl,et al.  Characteristics of Hodgkin's disease-derived cell lines. , 1982, Cancer treatment reports.

[7]  S. Masuda,et al.  A novel CD10-positive erythroid cell line, RM10, established from a patient with chronic myelogenous leukemia. , 1990, Leukemia.

[8]  H. Lazarus,et al.  Continuous culture of human lymphoblasts from peripheral blood of a child with acute leukemia , 1965, Cancer.

[9]  C. Tzeng,et al.  Establishment and characterization of a human monocytoid leukemia cell line, CTV-1. , 1984, Gan.

[10]  J. H. Brooke,et al.  Continuous tissue culture of leukocytes from human leukemic bloods by application of gradient principles. , 1955, Blood.

[11]  Richard Hurwitz,et al.  Characterization of a leukemic cell line of the pre‐B phenotype , 1979, International journal of cancer.

[12]  Hans G. Drexler,et al.  The Leukemia-Lymphoma Cell Line Factsbook , 2000 .

[13]  J. Masters.  HeLa cells 50 years on: the good, the bad and the ugly , 2002, Nature Reviews Cancer.

[14]  C. Fonatsch,et al.  Establishment and characterization of a novel CD34-positive human myeloid leukemia cell line: MHH225 growing in serum-free culture , 1995, Annals of Hematology.

[15]  Swee Lay Thein,et al.  Hypervariable ‘minisatellite’ regions in human DNA , 1985, Nature.

[16]  O. Markovic,et al.  Cell cross-contamination in cell cultures: The silent and neglected danger , 1998, In Vitro Cellular & Developmental Biology - Animal.

[17]  Y. Matsuo,et al.  Guidelines for the characterization and publication of human malignant hematopoietic cell lines , 1999, Leukemia.

[18]  Stephen J. O'Brien,et al.  Contamination of Hodgkin's disease cell cultures , 1981, Nature.

[19]  P. Koistinen,et al.  p53 status of newly established acute myeloid leukaemia cell lines , 1999, British Journal of Cancer.

[20]  H. Drexler,et al.  ECV304 (endothelial) is really T24 (bladder carcinoma): Cell line cross-contamination at source , 1999, In Vitro Cellular & Developmental Biology - Animal.

[21]  H. Nakauchi,et al.  Establishment and characterization of a human leukemic cell line with megakaryocytic features: dependency on granulocyte-macrophage colony-stimulating factor, interleukin 3, or erythropoietin for growth and survival. , 1991, Cancer research.

[22]  D. Gumucio,et al.  Granulocyte-macrophage colony-stimulating factor-dependent growth and erythropoietin-induced differentiation of a human cell line MB-02. , 1991, Blood.

[23]  A. Jeffreys,et al.  Individual-specific ‘fingerprints’ of human DNA , 1985, Nature.

[24]  H. Stein,et al.  Phenotypic analysis of an established cell line derived from a patient with Hodgkin's disease (HD) , 1985, Hematological oncology.

[25]  M. Greaves,et al.  Establishment and characterization of a new leukaemic T‐cell line (Peer) with an unusual phenotype , 1980, International journal of cancer.

[26]  G. Moore,et al.  Leukocyte cultures of patients with leukemia and lymphomas. , 1966, New York state journal of medicine.

[27]  S. Collins,et al.  Continuous growth and differentiation of human myeloid leukaemic cells in suspension culture , 1977, Nature.

[28]  C. Alvarado,et al.  Two new acute lymphoblastic leukemia cell lines with early B-cell phenotypes. , 1982, Blood.

[29]  M. Schmid,et al.  Digoxigenated oligonucleotide probes specific for simple repeats in DNA fingerprinting and hybridization in situ , 1989, Human Genetics.

[30]  R. Ádány,et al.  Marker profile, enzyme activity, and function of a human myelomonocytic leukemia cell line. , 1992, Cellular immunology.

[31]  M. Lilly,et al.  Characterization of a new human diploid myeloid leukemia cell line (PLB-985) with granulocytic and monocytic differentiating capacity. , 1987, Blood.

[32]  N. Harris.  Plasma cell surface antigen on human blood lymphocytes , 1974, Nature.

[33]  K. Nilsson,et al.  Establishment and characterization of a human histiocytic lymphoma cell line (U‐937) , 1976, International journal of cancer.

[34]  H. Drexler,et al.  Identity of original and late passage Dami megakaryocytes with HEL erythroleukemia cells shown by combined cytogenetics and DNA fingerprinting , 1997, Leukemia.

[35]  C. James,et al.  Homozygous deletions of the CDKN2 (MTS1/p16ink4) gene in cell lines established from children with acute lymphoblastic leukemia. , 1995, Leukemia.

[36]  H. Drexler,et al.  False human hematopoietic cell lines: cross-contaminations and misinterpretations , 1999, Leukemia.

[37]  R. Mohammad,et al.  Establishment of a human B-CLL xenograft model: utility as a preclinical therapeutic model. , 1996, Leukemia.

[38]  M. Theodoropoulou,et al.  Autocrine IL-2-dependent growth of a newly established CD3+, CD16−, CD56+, CD57+, JH−, TCRβ−, TCRγ− leukemia cell line (NOI-90) , 1997, Leukemia.

[39]  P. Marynen,et al.  A new subtype of pre-B acute lymphoblastic leukemia with t(5;12)(q31q33;p12), molecularly and cytogenetically distinct from t(5;12) in chronic myelomonocytic leukemia. , 1997, Blood.

[40]  L. Skinnider,et al.  Characteristics of a cell line established from a case of acute megakaryoblastic leukemia. , 1997, Acta Haematologica.

[41]  C. R. Barker,et al.  The establishment and cytological, cytochemical and immunological characterisation of human haemic cell lines: Evidence for heterogeneity , 1977 .

[42]  W. Schmidt,et al.  Hemoglobins from Plasmodium-infected rat erythrocytes: functional and molecular characteristics. , 1994, Blood.

[43]  J. Hoxie,et al.  Phenotypic and functional analysis of B cell lines from patients with multiple myeloma. , 1985, Blood.

[44]  H. Drexler,et al.  Authentication and Characterization , 2002 .

[45]  M. Minden,et al.  Expression of a retinoic acid receptor gene in myeloid leukemia cells. , 1989, Leukemia.

[46]  H. Drexler,et al.  Cross-contamination: HS-Sultan is not a myeloma but a Burkitt lymphoma cell line. , 2001, Blood.

[47]  S. Morikawa,et al.  Two E‐rosette‐forming lymphoid cell lines , 1978, International journal of cancer.

[48]  V. P. Collins,et al.  KBM-3, an in vitro model of human acute myelomonocytic leukemia. , 1992, Experimental hematology.

[49]  K. Mukai.  Large cell lymphoma. , 1989 .

[50]  P. Moorhead,et al.  Involvement of C Group Chromosomes in Five Burkitt Lymphoma Cell Lines , 1967 .

[51]  H. Drexler,et al.  Authentication of cancer cell lines by DNA fingerprinting. , 2004, Methods in molecular medicine.

[52]  D. Rosenthal,et al.  Characterization of a new megakaryocytic cell line: the Dami cell. , 1988, Blood.

[53]  K. Hirschhorn,et al.  Observations on cell lines derived from a patient with Hodgkin's disease. , 1978, Cancer research.

[54]  C. Lozzio,et al.  Cytotoxicity of a factor isolated from human spleen. , 1973, Journal of the National Cancer Institute.

[55]  G. Hodgson,et al.  Cell lines derived from a human myelomonocytic leukaemia , 1982, British journal of haematology.

[56]  H. Drexler,et al.  Persistent use of misidentified cell lines and its prevention , 2002, Genes, chromosomes & cancer.

[57]  H. Drexler,et al.  First searchable database for DNA profiles of human cell lines: sequential use of fingerprint techniques for authentication. , 1999, Cellular and molecular biology.

[58]  R. Handin.  DAMI cell line. , 1997, Blood.

[59]  R. Bataille,et al.  Human myeloma cell lines as a tool for studying the biology of multiple myeloma: a reappraisal 18 years after. , 1995, Blood.

[60]  T. Suda,et al.  Models of lineage switching in hematopoietic development: a new myeloid-committed eosinophil cell line (YJ) demonstrates trilineage potential , 1998, Leukemia.

[61]  T. Tanaka,et al.  Characterization, growth, and differentiation of a human myeloid leukemia cell line, TI-1 cell. , 1992, Blood.

[62]  J. Cossman,et al.  TPA-induced arrest of erythroid differentiation is coupled with downregulation of GATA-1 and upregulation of GATA-2 in an erythroid cell line SAM-1. , 1996, Blood.

[63]  J. Sklar,et al.  Establishment of cell lines from B-cell precursor acute lymphoblastic leukemia. , 1993, Leukemia.

[64]  G. Klein,et al.  Establishment and characterization of an Epstein-Barr virus (EBC)-negative lymphoblastoid B cell line (BJA-B) from an exceptional, EBV-genome-negative African Burkitt's lymphoma. , 1975, Biomedicine / [publiee pour l'A.A.I.C.I.G.].

[65]  R A F MacLeod,et al.  DNA profiling and cytogenetic analysis of cell line WSU-CLL reveal cross-contamination with cell line REH (pre B-ALL) , 2002, Leukemia.

[66]  H. Drexler,et al.  Differences in DNA fingerprints of continuous leukemia-lymphoma cell lines from different sources. , 1992, Leukemia.

[67]  M. Muto,et al.  Establishment and characterization of a novel human cell line exhibiting both immunophenotypic markers of monocyte/macrophage and natural killer cell lineages from peripheral blood of a patient with atopic dermatitis , 1997, Experimental dermatology.

[68]  G. Bornkamm,et al.  Characterization of EBV‐genome negative “null” and “T” cell lines derived from children with acute lymphoblastic leukemia and leukemic transformed non‐Hodgkin lymphoma , 1977, International journal of cancer.

[69]  A. Aventín,et al.  A new human cell line with pre-B cell phenotype and t(5;12) , 1997, Leukemia.

[70]  J. Fahey,et al.  PROLIFERATION AND DIFFERENTIATION OF LYMPHOID CELLS: STUDIES WITH HUMAN LYMPHOID CELL LINES AND IMMUNOGLOBULIN SYNTHESIS , 1971, Annals of the New York Academy of Sciences.

[71]  K. Inokuchi,et al.  p53 and N-ras mutations in two new leukemia cell lines established from a patient with multilineage CD7-positive acute leukemia. , 1993, Blood.

[72]  D. Horsman,et al.  Establishment and characterization of a human leukemic cell line (SR-91) with features suggestive of early hematopoietic progenitor cell origin. , 1994, Leukemia & lymphoma.

[73]  P. Nowell,et al.  Growth factor requirements of childhood acute leukemia: establishment of GM-CSF-dependent cell lines , 1987 .

[74]  W. Nelson-Rees,et al.  HeLa cultures defined. , 1976, Science.

[75]  R. E. Ritts,et al.  Establishment and characterization of a human non‐secretory plasmacytoid cell line and its hybridization with human B cells , 1983, International journal of cancer.

[76]  G. Stacey,et al.  Cell contamination leads to inaccurate data: we must take action now , 2000, Nature.

[77]  R. Cody,et al.  Characterization of a new human multiple myeloma cell line, UMJF-2, which suppresses antibody production by B-lymphocytes in vitro. , 1991, Leukemia.

[78]  D. Arthur,et al.  Establishment of a leukemic cell model for studying human pre-B to B cell differentiation. , 1989, Journal of immunology.

[79]  H G Drexler,et al.  HeLa cross-contamination of a leukemia cell line. , 1993, Leukemia.

[80]  M. Greaves,et al.  Phenotypic characterisation of a unique non-T, non-B acute lymphoblastic leukaemia cell line , 1977, Nature.

[81]  J. Minowada,et al.  ARH‐77, an established human IgG‐producing myeloma cell line. I. morphology, B‐cell phenotypic marker profile, and expression of epstein‐barr virus , 1984, Cancer.

[82]  H. Drexler,et al.  Malignant hematopoietic cell lines: in vitro models for the study of multiple myeloma and plasma cell leukemia. , 2000, Leukemia research.

[83]  John R. W. Masters,et al.  Human cancer cell lines: fact and fantasy , 2000, Nature Reviews Molecular Cell Biology.

[84]  V. Diehl,et al.  Hodgkin's disease: Establishment and characterization of four in vitro cell lines , 2004, Journal of Cancer Research and Clinical Oncology.

[85]  R. Huebner,et al.  Propagation of human tumors in antithymocyte serum-treated mice. , 1974, Journal of the National Cancer Institute.

[86]  J A Thomson,et al.  Short tandem repeat profiling provides an international reference standard for human cell lines , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[87]  G. Faguet,et al.  Cytochemical, cytogenetic, immunophenotypic and tumorigenic characterization of two hairy cell lines. , 1988, Blood.

[88]  R. Class,et al.  Cytokine-mediated erythroid maturation in megakaryoblastic human cell line HU-3. , 1997, Experimental hematology.

[89]  F. Elder,et al.  Role of marrow stromal cells in the establishment of a transformed lymphoblastic B-cell line from a normal human subject. , 1986, Leukemia research.

[90]  H. Drexler,et al.  Widespread intraspecies cross‐contamination of human tumor cell lines arising at source , 1999, International journal of cancer.

[91]  H. Drexler,et al.  Recent results on the biology of Hodgkin and Reed-Sternberg cells. II. Continuous cell lines. , 1993, Leukemia & lymphoma.

[92]  H. Drexler,et al.  Multiparameter approach in the identification of cross-contaminated leukemia cell lines. , 1993, Leukemia & lymphoma.

[93]  M. Taniwaki,et al.  Establishment of CD7+ Human Myeloma Sister Cell Lines, KMS-21-PE and KMS-21-BM, Carrying t(11;14) and t(8;14) , 2001, Leukemia & lymphoma.

[94]  H. Drexler,et al.  Cytogenetic characterization of tumor cell lines. , 2004, Methods in molecular medicine.

[95]  H. Drexler,et al.  Morphological, immunophenotypical and isoenzymatic profiles of human leukemia cells and derived T‐cell lines , 1989, Hematological oncology.

[96]  Y. Furukawa,et al.  Establishment and characterization of four human monocytoid leukemia cell lines (JOSK-I, -S, -M and -K) with capabilities of monocyte-macrophage lineage differentiation and constitutive production of interleukin 1. , 1986, Cancer Research.

[97]  W. Dai,et al.  HIMeg-1, a cell line derived from a CML patient, is capable of monocytic and megakaryocytic differentiation. , 1995, Leukemia.

[98]  A. Feller,et al.  Characterization of the novel, pediatric Hodgkin disease-derived cell line HKB-1. , 1998, Medical and pediatric oncology.

[99]  Donald Evans,et al.  Betrayers of the Truth: Fraud and Deceit in Science , 1986 .

[100]  H. Drexler,et al.  Mix-ups and mycoplasma: the enemies within. , 2002, Leukemia research.

[101]  Childhood lymphoblastic leukemia with natural killer activity: establishment of the leukemia cell lines retaining the activity , 1982 .

[102]  T. Abe,et al.  A novel human myelomonocytoid cell line, P39/Tsugane, derived from overt leukemia following myelodysplastic syndrome. , 1984, Gan.

[103]  L. Atkins,et al.  Tissue culture studies in Hodgkin’s disease morphologic, cytogenic, cell surface, and enzymatic properties of cultures derived from splenic tumors , 1977, The Journal of experimental medicine.

[104]  S. O’Brien,et al.  Application of DNA fingerprints for cell-line individualization. , 1990, American journal of human genetics.

[105]  A. Borkhardt,et al.  Recurrent chromosomal translocations and fusion genes in leukemia-lymphoma cell lines. , 1995, Leukemia.

[106]  E. Thompson,et al.  Establishment and characterization of a human myeloid cell line from Philadelphia chromosome-negative myeloblastic leukemia arising in a patient with myelodysplastic syndrome. , 1987, Blood.