Download Continuous and Primary Tissue Cell Line Cultures

1055 East State Street, Suite 100
Athens, OHIO 45701 USA
USA Toll-free: (800) 874-1517
International: +1(858) 552-1100
Fax: +1(740) 592-9820
www.dhiusa.com
QUALITY ASSURANCE STATEMENT
Continuous and Primary Tissue
Cell Line Cultures
Diagnostic Hybrids, Inc. (DHI) manufactures products in a facility certified to the ISO 13485:2003 quality
system standard for medical device manufacturers and in conformance with the requirements of USA’s
Code of Federal Regulations title 21 (CFR), the requirements of Canada’s Medical Device Regulations
(CMDR), and the requirements of European Union’s In Vitro Diagnostic Medical Devices Directive (IVDD).
DHI is the sole manufacturer of FreshCells™, MixedCells™, ReadyCells® and FreshFrozenCells® cell
culture products for in vitro diagnostic use.
 Both continuous cell lines and primary kidney tissue cultures are processed in separate, isolated and
distinct cleanroom areas. In addition, only one continuous cell line is processed at a time.
 Since cell cultures have a relatively short shelf-life, cells cannot be tested for the required 14 days prior
to shipment to verify absence of microorganisms. DHI continues to monitor representative samples
from each lot of cell product for evidence of microorganisms for at least 14 days after first shipment.
CONTINUOUS CELL LINES: A continuous cell line is a group of morphologically uniform cells that can be
propagated in vitro and are capable of an unlimited number of population doublings for an indefinite time.
Continuous cell lines usually either come from tumor tissue or have been deliberately immortalized or
transformed. Continuous cell cultures are intended for virus and/or Chlamydia isolation and to aid in the
diagnosis of diseases associated with infectious agents. These cells should not be used for serial cell
propagation.
 Cell types are from reliable, reputable, and traceable sources. Prior to acceptance into DHI’s
production facility, cell lines and strains are reviewed through documentation history and laboratory
analysis to verify that no microorganisms (by sterility testing) and no viruses [as evidenced by the
absence of cytopathic effect (CPE)] are known to be present.
 Frozen stock cultures are screened for the absence of Mycoplasma spp. and other adventitious
microorganisms.
 The various cell types are characterized as to species identity by isoenzyme analysis.
 Prior to shipment of each lot of cell culture, the culture medium is examined for clarity and pH, and
monolayers are examined microscopically for morphology, confluence, and uniformity.
 During a cell product’s lifetime, representatives of each batch are evaluated according to defined
performance criteria using appropriate indicator viruses or Chlamydia as inoculae.
Product
40-MC4- (CHO-Mc4)
43- (HEL)
44- (MRHF)
51- (MRC-5)
52- (CV-1)
53- (BGMK)
54- (McCoy)
55- (ELVIS® HSV)
56- (A549)
57- (Hep-2)
58- (Mv1Lu)
59- (NCI-H292)
62- (MNA)
67- (Vero 76)
Continuous Cell Culture Description
Chinese hamster ovary, genetically modified
human embryonic lung
human foreskin fibroblast
human embryonic lung fibroblast
African green monkey kidney
Buffalo green monkey kidney
mouse fibroblast
baby hamster kidney, genetically modified (BHK)
human lung carcinoma
human epidermoid carcinoma
mink lung epithelial
human pulmonary muco-epidemoid carcinoma
mouse neuorblastoma
African green monkey kidney
Product
70- (Super BGMK)
75- (HeLa 229)
76- (RD)
83- (MDCK)
84- (Vero)
85- (WI-38)
86- (LLC-MK2)
87- (Hs27 / HFF)
92- (Super E-Mix™)
96- (R-Mix™)
97- (R-Mix Too™)
98- (H&V-Mix™)
92- (Super E-Mix™)
Continuous Cell Culture Description
Buffalo green monkey kidney, genetically modified
human cervix adenocarcinoma
human rhabdomyosarcoma
Madin-Darby canine kidney
African green monkey kidney
human embryonic lung fibroblasts
Rhesus monkey kidney
human foreskin fibroblast
A549 and BGMK-hDAF
A549 and Mv1Lu
A549 and MDCK
MRC-5 and CV-1
A549 and BGMK-hDAF
PRIMARY KIDNEY TISSUE CULTURES: Primary Monkey Kidney (PMK) tissue cultures [cells isolated from the
kidneys of Macaca mulatta (Rhesus), Macaca fascicularis (Cynomolgus), or Cercopithecus aethiops (African
Green)] have been shown to be very permissive to human viruses and are intended to be used in the
cultivation and amplification of viruses, such as respiratory viruses, enteroviruses, measles, and poliovirus
from patient specimens. PMK cells are derived from adult animals by enzyme treatment of kidney tissues.
The cells are centrifuged free of the enzyme preparation, re-suspended in growth medium and cultured in
appropriate containers.
REF: PI-038.6en v2012JUN04
© 2012 Diagnostic Hybrids, Inc., Athens, OH 45701 USA.
1055 East State Street, Suite 100
Athens, OHIO 45701 USA
USA Toll-free: (800) 874-1517
International: +1(858) 552-1100
Fax: +1(740) 592-9820
www.dhiusa.com
QUALITY ASSURANCE STATEMENT
Continuous and Primary Tissue
Cell Line Cultures
PMK cells, like all primary animal cells, may harbor endogenous viruses1,2 such as simian viruses (SV), foamy
viruses, adenoviruses3, Herpes B virus, etc. as well as bacteria and parasites. These PMK culture
endogenous viruses can sometimes interfere with the culture of virus contained in the inoculum. It should be
noted that Herpes B, though innocuous to monkeys can infect humans and cause death. The most common
endogenous viruses are SV5 and SV40. Addition of antisera to SV5 and SV40 to the medium may inhibit the
growth of these SV and is, therefore, available as an option for the Rhesus products. Some of these viruses
may be expressed only after the cells have been “stressed” by, for example, environmental factors, age of the
culture, or inoculation with a specimen or fresh medium. Consequently, all results obtained using this product
should be reviewed by appropriately trained laboratory personnel, aware of the possibility that observed CPE
may be caused by an endogenous virus as well as a virus in the specimen.
Testing performed on primary kidney tissue by Supplier*
Product
Source
Herpes B virus
46- (pAGMK)
African Green
NO (not a carrier)
47- (pCMK)
Cynomolgus
YES - tested
48- (pRK)
Rabbit, New Zeal§
NO - not required
49- (pPhMK)
Rhesus
YES - tested
Other test or vaccinations
tuberculosis (TB) test; rabies, tetanus
tissue tested negative up removal, see below
closed pathogen free colony; 7-day sterility
tissue tested negative up removal, see below
*Generally, a brief life history and health certificate is provided with each monkey kidney (e.g., tracking code, source code,
animal ID number, age, physical traits, clinical record, vaccinations, treatment for parasites, etc.). Specific tissue tests performed
by each supplier varies; tissue used is negative using PCR or ELISA for: E. coli, Ebola, Hepatitis, Herpes B, Marburg, Measles,
Mycoplasma, SIV and SFV. The standard monkey colony practice is to vaccinate for rabies and tetanus, and test for TB.
§
Rabbit kidney tissue (New Zeal White) is supplied from a closed specific pathogen free colony; the kidney is aseptically
removed and placed in sterile transport medium; direct tests are performed on specific tissue to check for potential pathogens.
GENERAL CELL CULTURE PRECAUTIONS:
1. As with all methods for virus identification using cultured cells, personnel must be properly trained in virus
culture and safe handling techniques as described in the CDC-NIH manual2, Biosafety in Microbiological
and Biomedical Laboratories, i.e., manipulations which present potential personnel hazards should be
conducted in a Class II biosafety cabinet and gloves should be worn at all times. Cultured cells used for
virus identification may also support the replication of infectious agents which are classified by the CDC as
agents requiring cultivation under BSL-3 conditions. Consult CDC listing of the BSL-3 infectious agents and
the CDC recommendations.
2. The laboratory must determine the cell type to be used as host for isolation of a particular virus and/or
Chlamydia. Use for diagnostic procedures without prior culture has not been established.
3. Upon receipt of all cell cultures, the uninoculated monolayer should be examined for appearance (e.g., dead
or toxic cells, confluency) and morphology (e.g., atypical cells or CPE) prior to inoculation.
4. To aid in determining whether an endogenous virus might be present, negative controls (both with fresh
medium and as never-opened cultures) should be used and monitored for any cellular changes.
5. The classic detection method of viral infection is the observation of cellular changes due to infection of the
cells, termed cytopathic effect (CPE). CPE alone is not diagnostic and requires additional testing for viral
identification.
6. Some cell cultures have been shown to be non-permissive to infection and replication of the etiological
agents, such as, Severe Acute Respiratory Syndrome (SARS). For additional information, refer to the CDC
website at http://www.cdc.gov/ncidod/sars/lab/biosafety.htm.
7. Cultures and specimens should be autoclaved or disinfected with a solution of sodium hypochlorite (1:10
final dilution of household bleach) prior to disposal.
PRODUCT SPECIFICATIONS: Information beyond that provided by the Instructions for Use, Product Insert, Lot
Specification Report or Material Safety Data Sheet on a particular product or product lot may be request. DHI
will reviewed the information request and provide appropriate feedback.
2012 June 04
Karl E. Luke, PhD, Director, Quality Affairs
1
2
3
Date
G.D. Hsiung and V.F. Chan, “Endogenous viral and mycoplasma contaminants in cell cultures.”Hsiung’s Diagnostic
Virology, 4th ed; editors G.D. Hsiung, C.K.Y. Fong, and M.L. Landry; Yale Univ. Press; Ch 29, pp346-362, 1994.
CDC-NIH Manual, Biosafety in Microbiological and Biomedical Laboratories (BMBL), 5th edition, 2007, CDC-NIH manual.
[http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm]
Swierkosz, E.M. and T. Bonnot, Pseudoinfection due to Simian adenovirus, Clin. Microbiol. Newsletter, 14:14, 1992.
REF: PI-038.6en v2012JUN04
© 2012 Diagnostic Hybrids, Inc., Athens, OH 45701 USA.