Risk Assessment Approach to Microbiological Controls of Cell Therapies

Cundell, Tony; Drummond, Scott; Ford, Irving; Reber, Dona; Singer, Donald R. J.

doi:10.5731/pdajpst.2019.010546

Cited by 4 publications

(2 citation statements)

References 1 publication

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“…Microorganism contamination in the manufacturing of stem cells can occur through several sources, including raw materials, contaminated cell lines, operators, and processing (Mahmood and Ali, 2017;Cundell et al, 2020). Normally, to avoid the risk of microbial contamination (such as that from bacteria, yeast, and fungi) due to raw materials, the cell culture media are filtrationsterilized before use with sterilizing-grade filters of 0.2-0.22 μm nominal upper cut-off size.…”

Section: Introductionmentioning

confidence: 99%

“…In order to mitigate the risk of virus contamination for point-ofuse biopharmaceutical manufacturing, multi-barrier defence approaches are used, such as (i) working with low-risk starting and raw materials and using manufacturing controls; (ii) using multiple in-process controls for early detection of contamination and lot rejection; and (iii) preventive virus clearance measures to protect the cell lines from exposure to essential albeit high-risk raw materials (Cundell et al, 2020). Thus, the research gaps in point-ofuse biopharmaceutical manufacturing, specifically with regard to viral safety, lie in the need for integrated approaches involving testing, risk assessment, and prevention of contamination through physical barriers.…”

Section: Introductionmentioning

confidence: 99%

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Differentiation of human pluripotent stem cells into insulin-producing islet-like clusters using nanofiltered cell culture medium

Thorngren,

Vasylovska,

Blanc

et al. 2024

Front. Membr. Sci. Technol.

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The challenge of using patient-specific, autologous stem cell therapies in clinical settings is the need for advanced cell processing and expansion technologies. These include decentralized, small-scale manufacturing at the point of care in hospitals. The highest risk for contamination in cell-based therapy products comes from animal- and human-derived components such as serum, blood components, and growth factors. To mitigate the risk of adventitious microorganism contamination, preventive measures like size-exclusion virus removal filtration of cell media components can be employed. This article examines the impact of nanofiltration using nanocellulose-based virus clearance filter paper on the differentiation of human pluripotent stem cells into insulin-producing pancreatic islets (SC-islets). The cells were monitored for biomarkers using flow cytometry and immunohistochemistry along the 7-stage differentiation protocol. The produced SC-islets were evaluated functionally using low and high glucose stimulation under dynamic perifusion conditions. Pluripotent stem cells grown in culture media filtered through 20 nm cut-off nanocellulose filters showed similar expression of desired biomarkers at each stage compared to the control group. At the end of stage 7, SC-islets exhibited a rounded shape and strong expression of insulin, glucagon, and somatostatin in both the control and filtered media groups. The present study demonstrates that SC-islets differentiated with nanofiltered media were functional.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Differentiation of human pluripotent stem cells into insulin-producing islet-like clusters using nanofiltered cell culture medium

Thorngren,

Vasylovska,

Blanc

et al. 2024

Front. Membr. Sci. Technol.

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Comprehensive Study Identifies a Sensitive, Low-Risk, Closed-System Model for Detection of Fungal Contaminants in Cell and Gene Therapy Products

Putnam

Lau

2021

J Clin Microbiol

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The United States Food and Drug Administration (FDA) regulates manufacturing and testing of advanced therapeutic medicinal products (ATMPs) to ensure the safety of each product for human use. Gold standard sterility testing (USP<71>) and alternative blood culture systems have major limitations for the detection of fungal contaminants. In this study, we evaluated the performance of i LYM media (designed originally for the food and beverage industry) to assess its potential for use in the biopharmaceutical field for ATMP sterility testing. We conducted a parallel evaluation of four different test systems (USP<71>, BacT/ALERT, BACTEC, and Sabouraud Dextrose Agar [SDA] culture), three different bottle media formulations ( i LYM, i FA + , and Myco/F Lytic), and two incubation temperatures (22.5°C and 32.5-35°C) using a diverse set of fungi ( n =51) isolated from NIH cleanroom environments and previous product contaminants. Additionally, we evaluated the effect of agitation versus “delayed entry” static pre-incubation on test sensitivity and time to detection (TTD). Overall, delayed entry of bottles onto the BacT/ALERT or BACTEC instruments (with agitation) did not improve test performance. USP<71> and SDA culture continued to significantly outperform each automated culture condition alone. However, we show for the first time, that a closed-system, dual-bottle combination of i LYM 22.5°C and i FA + 32.5°C can provide high fungal sensitivity, statistically comparable to USP<71>, when tested against a diverse range of environmental fungi. Our study fills a much-needed gap in biopharmaceutical testing and offers a favorable testing algorithm that maximizes bacterial and fungal test sensitivity whilst minimizing risk of product contamination associated with laboratory handling.

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Report of a clinical and laboratory management of cell therapy for knee cartilage in the face of mycoplasma contamination

Zorzi¹,

Antonioli²,

Godoy³

et al. 2022

Einstein (São Paulo)

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To describe a case of autologous chondrocyte implantation after cell culture contamination by Mycoplasma pneumoniae and the measures taken to successfully complete cell therapy in a patient with focal chondral lesion. A 45-year-old male patient, complaining of chronic pain on the knee and no history of trauma. He had a chondral lesion in the trochlear region of the femur and clinical tests compatible with pain in the anterior compartment of the knee. Conservative treatment failed to alleviate symptoms. Surgical treatment was indicated, but due to the size of the lesion, membrane-assisted autologous chondrocyte implantation was the technique of choice. Cartilage biopsies were collected from the intercondylar region of the distal femur. After isolation, chondrocytes were expanded ex vivo in a trained laboratory, for three weeks, and seeded onto a commercially available collagen membrane prior to implantation in the knee. Two days before surgery, a cell culture sample tested positive for Mycoplasma pneumoniae. The source of contamination was found to be autologous blood serum, extracted from the patient´s peripheral vein, and used to supplement the cell culture medium. After treating the patient with antibiotics, all procedures were repeated and the new final cell product, free from contaminants, was successfully implanted. We discuss the strategies available to deal with this situation, and describe the results of this particular case, which led to modifications in the autologous chondrocyte implant protocol.

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Risk Assessment Approach to Microbiological Controls of Cell Therapies

Cited by 4 publications

References 1 publication

Differentiation of human pluripotent stem cells into insulin-producing islet-like clusters using nanofiltered cell culture medium

Differentiation of human pluripotent stem cells into insulin-producing islet-like clusters using nanofiltered cell culture medium

Comprehensive Study Identifies a Sensitive, Low-Risk, Closed-System Model for Detection of Fungal Contaminants in Cell and Gene Therapy Products

Report of a clinical and laboratory management of cell therapy for knee cartilage in the face of mycoplasma contamination

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