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Blood, 1 December 2001, Vol. 98, No. 12, pp. 3495-3496
CORRESPONDENCE
To the editor:
Cross-contamination: HS-Sultan is not a myeloma but a Burkitt
lymphoma cell line
Continuous hematopoietic cell lines have become important
research tools, contributing significantly to a better understanding of
the pathophysiology of hematopoietic tumors.1,2 A
relatively large number of cell lines have also been derived from
patients with multiple myeloma and related diseases such as plasma cell leukemia, plasmacytoma, etc.3,4 Unfortunately, a
high percentage of hematopoietic cell lines seem to have been
cross-contaminated with cells from other cell lines, which, as they
have a growth advantage, overgrew completely the original
culture.5 Cell line Jijoye was established from the tumor tissue of a 7-year-old
boy with Burkitt lymphoma in Africa in 1963.6,7 The cell
line HS-Sultan was allegedly established from the plasmacytoma of a
56-year-old Caucasian man who had IgG multiple myeloma.8 In a recent study by Davies et al (and also in a large number of
previous studies by this group), the continuous cell line HS-Sultan was
used as a "multiple myeloma" cell line.9 However, it
has been clearly established that this cell line is in reality a
subclone of the Burkitt lymphoma-derived cell line Jijoye and not a
myeloma cell line.3,8,10,11 The evidence is 5-fold: most
importantly DNA fingerprinting, but also viral, cytogenetic, immunologic, and morphologic findings. Due to insufficient
documentation of the allegedly "new" cell line and hence caused by
the lack of a proper karyotype of the original HS-Sultan cells, the
circumstances of the cross-contamination can no longer be traced back.
Most likely, during the attempts to establish a cell line, the cell culture was cross-contaminated with Jijoye cells, so there probably never was an "individual" HS-Sultan. DNA fingerprinting using different methodologic approaches at the
cell line banks American Type Culture Collection (ATCC) and the German
Collection of Microorganisms and Cell Cultures (German abbreviation
DSMZ) document independently and unequivocally that cell lines
HS-Sultan and Jijoye carry identical DNA fingerprints (Figure
1).
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| Figure 1.
High-resolution DNA fingerprint analysis of cell lines
Jijoye and HS-Sultan.
A multiple-locus DNA profile of the indicated cell lines was generated
using 10 µg of DNA digested to completion (HinfI),
size-separated on agarose gels and blotted on positively-charged nylon
membranes. After hybridization with a digoxigenin-labelled
(GTG)5 oligonucleotide as probe, a chemoluminescent
detection of (GTG)5 was carried out and the blot exposed
for 15 minutes to X-ray films. Molecular weight markers were used for
size determination. Cervix carcinoma cell line HELA was used as
positive control. The methods used have been described in details
elsewhere.11,13
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Both Jijoye and HS-Sultan are Epstein-Barr virus-positive
(EBV+).8 While all African-type Burkitt
lymphomas are EBV+ (including the cell lines derived from
these tumors), myeloma is clearly not an EBV-associated disease;
neither EBV DNA nor EBV nuclear antigen (EBNA) have been detected in
myeloma cells in the tumor. Myeloma cells are usually refractory to EBV
immortalization. There is no bona fide EBV+ myeloma cell
line.4 Our cytogenetic analysis showed that both Jijoye and HS-Sultan have a
similar though not identical hyperdiploid karyotype including
t(8;14)(q24;q32), occurring in 75%-85% of all Burkitt lymphoma
cases,12 together with partial or complete
trisomy of chromosome 7. Their karyotypes are as follows: Jijoye,
46/47 2n X/XY,dup(7)(q35q36),t(8;14)(q24;q32),der(14)t(9;14)(q11;q32),  18,+2mar;
and HS-Sultan, 482nXY,+7,t(8;14)(q24;q32), +mar. EBV
is, of course, a polyclonal activator and any karyotypic differences may be either in vitro artefacts or attributable to the known propensity of Jijoye to give rise to phenotypically distinct subclones or a combination of both. It should be mentioned that the
t(8;14)(q24;q32) is not specific for Burkitt lymphoma and was also
found in some 6% of the bona fide myeloma cell lines.4 A comparative immunophenotyping analysis of myeloma and nonmyeloma cell
lines showed that HS-Sultan does not express a immunomarker profile
compatible with that of canonical myeloma cell lines.4,10 Finally, on May-Grünwald-Giemsa-stained cytospin slides,
HS-Sultan cells do not display the morphologic features that are typical of myeloma cell lines (deep basophilic cytoplasm, excentrically located nucleus, perinuclear clear hof, dense or clumped chromatin). On
the contrary, HS-Sultan cells show an abundance of vacuolization (typical for B-acute lymphoblastic leukemia and Burkitt lymphoma, also
described as FAB L3), large nuclei and prominent nucleoli. Cross-contamination of cell lines is a wide-spread problem and
invalidates many research results.13 DNA fingerprinting, which is the method of choice, has documented that about 15%-20% of
the currently "circulating" cell lines are cross-contaminated and
overgrown by other cells.5,11 Unfortunately, cell line HS-Sultan had been used indiscriminately in a large number of studies
(even after the true nature of this cell line had become known),
rendering some of the conclusions irrelevant to the biology of multiple
myeloma.10 Unless Anderson et al and other
investigators wish to assign Burkitt lymphoma to the entity of multiple
myeloma and related diseases, cell line HS-Sultan should not be
employed any longer as a model system for multiple myeloma.
Hans G. Drexler, Roderick A. F. MacLeod, and Willy G. Dirks
Correspondence: Hans G. Drexler, DSMZ-German Collection of
Microorganisms and Cell Cultures, Mascheroder Weg 1 B, D-38124
Braunschweig, Germany; e-mail: hdr{at}dsmz.de
References
1.
Drexler HG.
The leukemia-lymphoma cell line facts book. San Diego, CA: Academic Press; 2000.
2.
Drexler HG, Matsuo Y, MacLeod RAF.
Continuous hematopoietic cell lines as model systems for leukemia-lymphoma research.
Leukemia Res.
2000;24:881-911[CrossRef][Medline]
[Order article via Infotrieve].
3.
Jernberg-Wiklund H, Nilsson K.
Multiple myeloma cell lines. In:
Masters JRW,Palsson BO, eds.
Human cell culture Vol 3. Cancer cell lines Part 3: Leukemias and lymphomas. Dordrecht, Netherlands: Kluwer; 2000:81-155.
4.
Drexler HG, Matsuo Y.
Malignant hematopoietic cell lines: in vitro models for the study of multiple myeloma and plasma cell leukemia.
Leukemia Res.
2000;24:681-703[CrossRef][Medline]
[Order article via Infotrieve].
5. Drexler HG, Uphoff CC, Dirks WG, MacLeod RAF. Mix-ups and mycoplasma:
the enemies within. Leukemia Res. 2001. In press.
6.
Pulvertaft RJV.
Cytology of Burkitt's tumour (African Lymphoma).
Lancet.
1964;1:238-240.
7.
Drexler HG, Minowada J.
History and classification of human leukemia-lymphoma cell lines.
Leukemia Lymphoma.
1998;31:305-316.
8. American Type Culture Collection (ATCC). Cell biology collection
catalog. Available at: www.atcc.org/searchcatalogs/cellbiology.cfm.
Accessed August 1, 2001.
9.
Davies FE, Raje N, Hideshima T, et al.
Thalidomide and immunomodulatory derivatives augment natural killer cell cytotoxicity in multiple myeloma.
Blood.
2001;98:210-216[Abstract/Free Full Text].
10.
Pellat-Deceunynk C, Amiot M, Bataille R, et al.
Human myeloma cell lines as a tool for studying the biology of multiple myeloma: a reappraisal 18 years after.
Blood.
1995;86:4001-4002[Free Full Text].
11.
Drexler HG, Dirks WG, MacLeod RAF.
False human hematopoietic cell lines: cross-contaminations and misinterpretations.
Leukemia.
1999;13:1601-1607[CrossRef][Medline]
[Order article via Infotrieve].
12.
Mitelman F.
Catalog of chromosome aberrations. 5th ed. New York, NY: Wiley-Liss; 1994.
13.
MacLeod RAF, Dirks WG, Kaufmann M, Matsuo Y, Milch H, Drexler HG.
Widespread intraspecies cross-contamination of human tumor cell line arising at source.
Int J Cancer.
1999;83:555-563[CrossRef][Medline]
[Order article via Infotrieve].
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