100 for 100: Looking back at ATCC’s most influential deposits

Item 9/100: The Poliovirus
Sarah Vannozzi, MS, Strategy Realization Principal, and Kati Kiss, PhD, Manufacturing Principal
ATCC® VR-59™ – Poliomyelitis, Type 1

The poliovirus was first isolated in 1909 by Austrian physicians Karl Landsteiner and Erwin Popper at the Wilhelminenspital in Vienna.¹ In 1955, Jonas Salk developed the inactivated poliovirus vaccine (IPV) using three strains of the “poliovirus”: Mahoney (type 1; ATCC® VR-59™), MEF-1 (type 2; an example is ATCC® VR-61™), and Saukett (type 3; an example is ATCC® VR-62™).² Originally during vaccine development, the viruses were propagated on monkey kidney cells; later, the monkey cells were replaced by HeLa cells (ATCC® CCL-2™) that were mass-produced at Tuskegee University to keep pace with the production needed to distribute the vaccine widely.³ Since the development of the vaccine, the type 2 wild poliovirus was declared eradicated in September 2015 with the last known detected case in India in 1999. The type 3 wild poliovirus was declared eradicated in October 2019; it was last detected in November 2012. Only the type 1 wild poliovirus remains.⁴

Why the polio vaccine is important to Sarah: “My dad was one of the 50 or so people who contracted polio in the summer of 1952 in Maine before the vaccine was developed and distributed. I am thankful that using these resources to develop vaccines means this disease has mostly been eradicated.”

Item 10/100: From Park to PCR
Kati Kiss, PhD, Manufacturing Principal and Brian Chase, MS, Supervisor Laboratory Operations
ATCC® 25104™ – Thermus aquaticus Brock and Freeze

In the late 1960s, Thomas Brock and Hudson Freeze were collecting samples from the hot springs in Yellowstone National Park. At the time, thermophilic bacteria had been described in the literature but were limited to spore-forming species. In 1966, a non-sporulating, gram-negative rod was isolated from a sample taken from the 71.5°C (160°F) Mushroom Spring in the Lower Geyser Basin of the park. The organism was later named Thermus aquaticus YT-1 and was deposited with ATCC in 1968.⁵

By the mid-1980s, polymerase chain reaction (PCR) was a technique that was being used in laboratories to amplify DNA. In the early days of this technique, DNA polymerase from Escherichia coli was used to generate the new daughter strands; however, this enzyme is heat sensitive and needed to be refreshed though out the process. Finding a heat stable polymerase for the process was therefore critical, but purifying the heat stable polymerase from thermophiles was not easy.⁶ This all changed when the polymerase gene from Thermus aquaticus strain YT-1 (ATCC® 25104™) was cloned into E. coli to generate the recombinant polymerase Taq Pol I. This enzyme was introduced to the molecular biology world via a publication by RK Saiki et al. in 1988, initiating a biotechnology revolution with impacts spanning disease research to forensics.⁷

As part of ATCC’s mission, we continue to support deposits of microbial species found in US national parks within the US National Park Service Special Collection. This unique collection of unusual microbes is available to support future advancements in science, with over 100 species and growing.

Item 11/100: The CHO-sen One
Ana Eckert, BS, Senior Innovation Specialist
ATCC® CCL-61™ – CHO-K1

Unbeknownst to most, today’s world of modern medicine relies heavily on Chinese hamster ovarian (CHO) cells. These workhorse cells readily survive in bioprocessing environments because they are extremely tolerant of variations in pH, oxygen levels, temperature, and pressure, among other variables.⁸ When researchers discovered in the 1950s that CHO cells could multiply rapidly and produce large amounts of protein, they quickly became critical in the development of protein-based therapeutics. These therapeutics have been instrumental in treating a wide array of medical conditions, including cancer, hemophilia, psoriasis, hormone imbalances, and more. Today, scientists use gene-editing tools to engineer CHO cells to produce specific therapeutic antibodies or proteins—this has contributed to the production of most of the biologic drugs in the US and over half of the therapeutic antibodies used in cancer treatments.⁹

Item 12/100: Streptococcus pyogenes and the Development of CRISPR-Cas9
Scott Nguyen, PhD, Senior Biocuration Scientist
ATCC® 700294™ – Streptococcus pyogenes Rosenbach

Streptococcus pyogenes strain SF370 (ATCC® 700294™) has been instrumental in biotechnology.^10-12 In 2005, a study by Mojica et al. showed that short-repeated DNA sequences in ATCC® 700294™ were derived from mobile genetic elements.¹³ These repetitive genomic loci, also known as clustered regularly interspaced short palindromic repeats (CRISPR), were shown to confer immunity to foreign extrachromosomal elements. Later in 2012, the CRISPR-Cas9 endonuclease gene, cloned from ATCC® 700294™, was used in groundbreaking research by Jennifer Doudna and Emmanuelle Charpentier for programmable gene editing.¹⁴ The use of CRISPR-Cas9 for genome editing has revolutionized the biotechnology field and ultimately led to the Nobel Prize in Chemistry for Drs. Jennifer Doudna and Emmanuel Charpentier.¹⁵

Why Streptococcus pyogenes strain SF370 is important to Scott: “It was my doctoral studies mentor, Dr. W. Michael McShan, who deposited this Group A Streptococcus strain into the ATCC collection in 1997. My mentor was part of a sequencing team that generated the first complete genome of S. pyogenes in 2001,¹⁶ and the genome of ATCC® 700294™ was one of the first complete bacterial genomes published. The complete genome of this strain laid the foundations for one of the most significant genetic tools of the 21^st century. Thanks to the foresight of Dr. McShan and his colleagues, the deposition of this strain into ATCC and the resulting availability of this organism has helped shape molecular biology in the last decade.”

Item 13/100: The Legendary NIH/3T3 Fibroblast Cell Line
Paul Lovell, PhD, Associate Scientist
ATCC® CRL-1658™ – NIH/3T3 cell line and ATCC® CRL-1658.2™ – NIH/3T3.2

The mouse cell line NIH/3T3 (ATCC® CRL-1658™) is a spontaneously immortalized fibroblast cell line. NIH/3T3 cells were developed by George Todaro and Howard Green in 1962 and is one of the most widely utilized cell lines in general cell biology, cancer research, and transfection studies.¹⁷ These cells were originally known for their strong contact inhibition and are widely used in assays to determine a gene of interest’s potential to become oncogenic.¹⁸ Introduction of an overexpressed gene in NIH/3T3 cells may promote morphological changes that allows the cells to create dense foci areas. Over time, the NIH/3T3 cells have emerged as a multi-clonal cell population resulting in cell transformation and the loss of contact inhibition. ATCC recently restored this property through single cell cloning, generating a clonal derivative—NIH/3T3.2 (ATCC® CRL-1658.2™)—that maintains the original contact inhibition property associated with these cells. ATCC is currently the only biorepository that has NIH/3T3 cells with the restored contact inhibition property within their collection.

Item 14/100: Mould Juice
Kendra Grosso, BS, Supervisor Laboratory Operations
ATCC® 9478™ – Penicillium rubens Biourge

For Sir Alexander Fleming, described by his fellow researchers as “untidy,” finding unusual growth in his bacterial cultures was a regular occurrence. While working with Staphylococcus aureus in the fall of 1928, he noticed that one particular fungal contaminant appeared to have a lytic effect on the surrounding bacteria.¹⁹ Through controlled experiments with this mold, identified by his colleague Charles J. La Touche as Penicillium rubens,²⁰ he proved the inhibitory effect against medically relevant infectious bacteria. While initially referring to the substance as “mould juice,” he later named the compound penicillin. Sir Flemming published his results to little fanfare and less acceptance among the medical community. It was only after the widespread use of sulfonamides to treat infection and the introduction of concentrated penicillin by Sir Ernst Chain and Howard Florey in 1940 that the medical community finally took notice.

In May 1944, ATCC acquired the so-called “Fleming strain” of Penicillium from the Northern Regional Research Laboratory (NRRL) of the USDA.²¹ Over the last 80 years, the use of antimicrobial compounds has become the primary treatment for bacterial infections worldwide. Against the backdrop of growing antibacterial resistance, this strain continues to serve an important role in the development of new treatments for bacterial infections.

Item 15/100: A Reference Material for Gene Therapy Potency Testing
Kyle Young, MBA, Product Manager
ATCC® VR-1516™ – Human adenovirus 5

Safety is a primary concern for gene therapy development. Following the tragic death of Jesse Gelsinger in 1999,²² the Adenovirus Reference Material Working Group (ARMWG) was formed to provide resources for accurate measurement of the potency of injectable products to biotechnology and pharmaceutical firms working on adenovirus-based gene therapies.²³ With support from the US FDA and an international coalition of organizations, the Adenovirus Reference Material (ARM)— a purified, well-characterized preparation of Adenovirus 5 derived from ATCC catalog number VR-5™—was developed and manufactured.²² As a member of the ARMWG and the steward for distribution of ARM, ATCC has maintained it as catalog number VR-1516™ since its release in 2001. Demand grew in concert with gene therapy development leading to a depletion of inventory in late 2021. With few industry resources available to support another working group effort, ATCC has taken responsibility for ARM replenishment.

Item 16/100: The Deadliest Bacteria Known to Mankind
Joseph Thiriot, PhD, Manager, Laboratory Operations
ATCC® 27294™ – Mycobacterium tuberculosis TMC 102 [H37Rv]

In 1905, a sick patient in the Trudeau Sanatorium in Saranac, New York, was treated by Dr. Edward R. Baldwin. He took a sample from the patient and identified it as Mycobacterium tuberculosis.²⁴ Known by many names throughout history, such as ‘consumption’, the disease caused by this bacterium is tenaciously difficult to treat and rid from the body. M. tuberculosis is a master of stealth and subterfuge, using the body’s own defenses against itself to create walled blockades in the lungs called granulomas, which stop an otherwise effective immune response. M. tuberculosis has historically been the leading cause of death worldwide from an infectious disease and was recently temporarily passed by the SARS-CoV-2 pandemic.²⁵ The Trudeau Institute provided this same strain to ATCC in 1972. ATCC has stored this important isolate for over 55 years, providing this reference standard to institutions around the world for research and development purposes. In so doing, ATCC is actively participating in the fight against the top infectious disease worldwide.

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On average, ATCC adds 150 new, and potentially influential, items to its collection every year.

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References

1. Paul JR. A History of Poliomyelitis. (Yale studies in the history of science and medicine). New Haven, Conn: Yale University Press, 1971. ISBN 978-0-300-01324-5.
2. Kew OM, et al. Vaccine-derived polioviruses and the endgame strategy for global polio eradication. Annu Rev Microbiol 59: 587–635, 2005. PubMed: 16153180.
3. Turner T. Development of the polio vaccine: a historical perspective of Tuskegee University's role in mass production and distribution of HeLa cells. J Health Care Poor Underserved 23(4 Suppl): 5–10, 2012. PubMed: 23124495.
4. Global Polio Eradication Initiative. GPEI Strategy 2022-2026 [PDF], July 2023.
5. Brock TD, Freeze H. Thermus aquaticus gen. n. and sp. n., a nonsporulating extreme thermophile. J Bacteriol 98(1): 289-297, 1969. PubMed: 5781580.
6. Saiki RK, et al. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239(4839): 487-491, 1988. PubMed: 2448875.
7. Lawyer FC, et al. Isolation, characterization, and expression in Escherichia coli of the DNA polymerase gene from Thermus aquaticus. J Biol Chem 264(11): 6427-6437, 1989. PubMed: 2649500.
8. Jerabek T, et al. Life at the periphery: what makes CHO cells survival talents. Appl Microbiol Biotechnol 106(18): 6157–6167, 2022. PubMed: 36038753.
9. NIST. Cho and tell. Published December 21, 2018. Accessed January 2025. <https://www.nist.gov/news-events/news/2018/02/cho-and-tell>
10. McShan WM, McCullor KA, Nguyen SV. The Bacteriophages of Streptococcus pyogenes. Microbiol Spectr 7(3): 10.1128, 2019. PubMed: 31111820.
11. Nguyen SV, McShan WM. Chromosomal islands of Streptococcus pyogenes and related streptococci: molecular switches for survival and virulence. Front Cell Infect Microbiol 4: 109, 2014. PubMed: 25161960.
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13. Mojica FJM. Intervening Sequences of Regularly Spaced Prokaryotic Repeats Derive from Foreign Genetic Elements. J Mol Evol 60(2): 174–182, 2005. PubMed: 15791728.
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17. Todaro GJ, Green H. Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J Cell Biol 17(2): 299-313, 1963. PubMed: 13985244.
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20. Allison VD. Personal Recollections of Sir Almroth Wright and Sir Alexander Fleming. Ulster Med J 43(2): 89-98, 1974. PubMed: 4612919.
21. ATCC historical records
22. Hutchins B, et al. Working toward an adenoviral vector testing standard. Mol Ther 2(6): 532–534, 2000. PubMed: 11124052.
23. Hutchins B, et al. Development of a reference material for characterizing adenovirus vectors. Bioprocess J 1 (1): 25-28, 2002.
24. Steenken W, Oatway WH, Petroff SA. Biological studies of the tubercle bacillus: III. Dissociation and pathogenicity of the R. and S. variants of the human tubercle bacillus (H(₃₇)). J Exp Med 60(4): 515–540, 1934. PubMed: 19870319.
25. World Health Organization. Global Tuberculosis Report 2024. Geneva: World Health Organization; 2024. Accessed January 2025 <https://www.who.int/teams/global-tuberculosis-programme/tb-reports/global-tuberculosis-report-2024>