Primocin®

Antimicrobial agent for primary cells

Primocin® is designed to offer complete protection of primary cells from microbial contamination. Primocin® is a broad-spectrum antibiotic formulation that is gentle on your cells but lethal to the microbes. There are a number of common sources of microbial contamination especially in the isolation of primary cells from both animal and human tissue, including commensal flora and/or subclinical infections.

Use of Primocin® in primary cell cultures

Key Features:

• Active against bacteria, mycoplasma, and fungi
• A common addition to various standardized primary cell culture protocols
• Used throughout culturing process (e.g. culture media, wash solutions, and  biopsy storage media) 
• Highly cited in the literature

Primocin® is composed of four compounds, of which three, act directly on Gram-positive bacteria, Gram-negative bacteria, and mycoplasmas. The primary mode of action for these compounds is to block microbial DNA and protein synthesis. The fourth compound in Primocin® eradicates fungi, including yeasts, by directly targeting processes at the fungal membrane.

Primocin® is successfully used in many types of primary cell cultures (see citations) including:

• Differentiated mouse- and human-derived cells such as astrocytes and immune cells (e.g. natural killer cells)
• Stem cells such as mouse- and human-derived embryonic and pluripotent stem cells
• 3D cellular models (i.e. organoid and spheroids) such as colon epithelial and carcinoma organoids

Primocin® is made in-house at InvivoGen and is of the highest quality with rigorous quality control performed to ensure the absence of lot-to-lot variation.

Note: Primocin® is a trademark of InvivoGen registered with the United States Patent and Trademark Office.

Figures

InvivoGen recommends using Primocin® at 100 μg/ml, which represents a 1:500 dilution of stock solution (see table).

Specifications

Product concentration: 50 mg/ml

Recommended working concentration: 100 μg/ml

• 1 ml vial of Primocin® is sufficient for 500 ml of culture.
• 20 ml bottle of Primocin® is sufficient for 10 liters of culture.

Quality Control:

• Physicochemical characterization (pH and appearance) has been confirmed
• Endotoxin levels: < 0.5 EU/mg
• Potency has been validated on bacterial and fungal reference strains

Contents

Primocin® is provided as a cell culture tested, sterile filtered, light yellow solution at a concentration of 50 mg/ml.

This product is available in three pack sizes:

• ant-pm-05 - 5 x 1 ml vial (250 mg) 
• ant-pm-1 - 10 x 1 ml vial (500 mg)
• ant-pm-2 - 1 x 20 ml bottle (1 g)

Primocin® is shipped at room temperature.
Upon receipt, Primocin® can be stored at 4°C for 6 months or at -20°C for long-term storage

Details

Use of Primocin® in primary cell cultures

Primocin® is frequently cited in the literature for use in the protection of a number of different primary cell cultures. This includes (but is not limited to):

• Numerous human and murine primary cell cultures, including fibroblasts [1], glial cells [2], astrocytes [3], and immune cells (e.g. NK cells) [4].
• Culturing and reprogramming of human and murine embryonic cells [5] and pluripotent stem cells [6].
• 3D cell cultures including colon epithelial and carcinoma organoids [7] as well as breast, bladder, and prostate cancer organoids [8-10].
• A number of published standardized protocols such as tumor organoid-T-cell co-culture systems [11].

References:

1. Ferrer-Mayorga, G. et al., 2019. Vitamin D and Wnt3A have additive and partially overlapping modulatory effects on gene expression and phenotype in human colon fibroblasts. Sci Rep 9:8085.
2. Bussian T.J. et al., 2018. Clearance of senescent glial cells prevents tau-dependent pathology and cognitive decline. Nature. 562:578‑82.
3. Grabner, G.F. et al., 2016. Deletion of Monoglyceride Lipase in Astrocytes Attenuates Lipopolysaccharide-induced Neuroinflammation. J Biol Chem 291;913‑23.
4. Garcia-Beltran, W.F. et al., 2016. Open conformers of HLA-F are high-affinity ligands of the activating NK-cell receptor KIR3DS1. Nat Immunol 17:1067-74.
5. Kogata N. et al., 2018. Sox9 regulates cell state and activity of embryonic mouse mammary progenitor cells. Commun Biol. 1:228.
6. Park S. et al., 2018. Generation of human-induced pluripotent stem cells using a defined, feeder-free reprogramming system. Curr Protoc Stem Cell Biol 45.
7. Urbischek M. et al., 2019. Organoid culture media formulated with growth factors of defined cellular activity. Sci Rep 9:6193.
8. Sachs N. et al., 2018. A living biobank of breast cancer organoids captures disease heterogeneity. Cell. 172:373-86.
9. Lee S.H. et al., 2018. Tumor evolution and drug response in patient-derived organoid models of bladder cancer. Cell 173:515-28.
10. Xu H. et al., 2018. Organoid technology and applications in cancer research. J Hematol Oncol 11:116.
11. Cattaneo, C.M. et al., 2020. Tumor organoid–T-cell coculture systems. Nat Prot 15, 15-39.