ITS Animal-Free™ – Blood-free cell culture media supplement

Product Description

ITS Animal-Free™ (ITS AF) is a 100x liquid recombinant cell culture supplement containing insulin, transferrin, and selenium — the 3-component defined supplementation profile required for serum-free and chemically defined mammalian cell culture workflows. The formulation delivers 1.00 g/L insulin, 0.55 g/L transferrin, and 0.67 mg/L selenium in a chemically defined, animal-origin-free profile suitable for a broad range of biopharmaceutical, cell therapy, regenerative medicine, and research cell culture applications.

ITS Animal-Free is the foundational supplement in InVitria's ITS product family, providing the three core defined factors — recombinant human insulin for growth and glucose uptake signaling, recombinant human transferrin for iron delivery via the TFR1 receptor-mediated pathway, and selenium as sodium selenite for antioxidant enzyme function — without the ethanolamine or albumin additions found in more comprehensive ITSE and ITSE+A formulations. The 3-component format is appropriate for cell culture workflows where ethanolamine supplementation is provided separately, is not required for the cell type in use, or where a minimal defined supplement footprint is preferred. Unlike plasma-derived ITS supplements that rely on human donor plasma transferrin — introducing pooled donor variability, adventitious agent qualification requirements, and supply chain exposure to global plasma markets — ITS Animal-Free™ is formulated with recombinant human transferrin, eliminating plasma-derived raw materials and supporting ICH Q5A-aligned regulatory pathways.

ITS Animal-Free has been used in peer-reviewed clinical-stage research including cell therapy manufacturing applications. Hirayama et al. (2025) used ITS Animal-Free™ in the corneal endothelial cell differentiation medium supporting CLS001, a clinical-grade iPSC-derived cell therapy product transplanted into a human patient in a first-in-human study of bullous keratopathy, published in Cell Reports Medicine [DOI: pending confirmation from ITS AF source]. This establishes ITS AF as a cell culture supplement with demonstrated use in regulated, human-grade cell therapy manufacturing — not just research-scale applications. ITS Animal-Free™ cross-references with ITSE Animal-Free™ (4-component with ethanolamine) and ITSE+A™ (5-component with ethanolamine and rHSA) within InVitria's ITS product family, providing a range of defined supplementation options for serum-free and chemically defined media development. ITS Animal-Free™ is manufactured under an ISO 9001:2015-certified, cGMP-compliant quality management system and is available in [packaging formats — need specs]. Full regulatory documentation — including Certificate of Analysis, Safety Data Sheet, Animal-Free Statement, and cGMP documentation — is available on request.

Advantages

Validated in Clinical-Stage Cell Therapy Manufacturing

• ITS Animal-Free was used in the corneal endothelial differentiation medium for CLS001, a clinical-grade iPSC-derived cell therapy product transplanted into a human patient in a first-in-human study of bullous keratopathy (Hirayama et al., 2025, Cell Reports Medicine).
• This establishes ITS AF as a cell culture supplement with demonstrated use in regulated, human-grade cell therapy manufacturing — not just research-scale applications.

Defined, 3-Component Formulation with No Undefined Additives

• Fully defined composition: recombinant human insulin (1.0 g/L), recombinant human transferrin (0.55 g/L), and sodium selenite (0.67 mg/L). No carrier proteins, no stabilizers, no preservatives, no undefined components.
• No animal-derived components in any raw material, processing aid, manufacturing equipment, or packaging — supporting ICH Q5A compliance and simplifying TSE/BSE qualification for regulatory submissions.
• Simpler formulation than ITSE or ITSE+A by design — fewer variables to qualify, fewer inputs to document, and a cleaner starting point for process development teams building serum-free media.

Regulatory-Ready Documentation and Manufacturing

• Formulated and quality-controlled at InVitria's ISO 9001:2015-certified, cGMP-compliant facility in Junction City, Kansas.
• Complete regulatory package available: Certificate of Analysis, Safety Data Sheet, Guidelines for Use, Certificate of Origin and Animal-Free / TSE-BSE Statement, ISO 9001 certificate, and cGMP Statement.
• Endotoxin controlled to ≤ 1.0 EU/mL, tested and released on every lot, with full traceability from raw material sourcing through finished product.

Broad Mammalian Cell Compatibility

• Validated across CHO, HEK293, VERO, hybridoma, fibroblast, keratinocyte, primary cells (T cells, NK cells), and stem cells (iPSC, MSC, HSC, CD34+).
• Supports biopharmaceutical production, viral vector manufacturing (AAV, lentivirus), antibody production, and regenerative medicine workflows.
• Typical working concentration of 1x (1:100 dilution from the 100x concentrate), with 2–3x recommended for demanding cell types — dosing guidance that applies across the full range of supported applications.

Applications

Stem Cell Expansion & iPSC Maintenance ITS AF has been validated in iPSC expansion with maintained pluripotency marker expression and is used in serum-free media formulations for MSC, HSC, and CD34+ hematopoietic stem cell culture (see ITS AF iPSC expansion application note). The defined recombinant insulin–transferrin–selenium composition provides the essential non-serum inputs these cell types require for proliferation and metabolic stability. For iPSC or MSC applications where additional protein stabilization is required — particularly in long-term maintenance or sensitive differentiation workflows — ITSE+A™ adds recombinant human albumin to provide broader serum-function replacement.

Cell Therapy Manufacturing & iPSC Differentiation ITS Animal-Free has been validated in clinical-stage cell therapy manufacturing, including use as a component of the corneal endothelial differentiation medium for CLS001, an iPSC-derived cell therapy product transplanted into a human patient in a first-in-human study of bullous keratopathy (Hirayama et al., 2025, Cell Reports Medicine). The chemically defined, recombinant composition makes ITS AF particularly well suited to cell therapy workflows where regulatory traceability, raw material qualification, and adventitious-agent risk mitigation are central to CMC strategy. ITS AF has been used across iPSC, MSC, HSC, CD34+, and primary T and NK cell expansion and differentiation protocols (see ITS AF iPSC expansion application note). For cell therapy applications requiring additional protein stabilization in long-term maintenance or sensitive differentiation workflows, ITSE+A™ adds recombinant human albumin to the ITS backbone.

Biopharmaceutical & Recombinant Protein Production ITS AF supports serum-free and reduced-serum biomanufacturing across CHO, HEK293, hybridoma, and other production cell lines used in monoclonal antibody, recombinant protein, and viral vector workflows — including AAV and lentivirus production for gene therapy manufacturing. Recombinant insulin supports glucose uptake and metabolic signaling, InVitria's recombinant transferrin delivers iron via TFR1-mediated endocytosis without the donor variability or adventitious-agent risk of plasma-derived transferrin, and selenium supports glutathione peroxidase activity to protect cells from oxidative stress. The fully animal-free composition supports ICH Q5A compliance and simplifies TSE/BSE qualification for IND, BLA, and supplier qualification submissions. Lot-to-lot consistency across recombinant components reduces raw material variability in production workflows where reproducibility directly impacts titer and yield.

Vaccine & VERO Cell Manufacturing ITS AF is compatible with VERO cell culture workflows used in vaccine and viral production, providing the core ITS supplementation needed to support VERO cell proliferation and productivity in serum-free media. The animal-free, recombinant composition addresses the regulatory pressure from FDA, EMA, and PMDA to eliminate animal- and serum-derived components from vaccine manufacturing processes. For fully defined, complete VERO cell culture, ITS AF is compatible with InVitria's OptiVERO® — a chemically defined complete medium designed specifically for VERO-based vaccine manufacturing without serum or animal-derived components.

Research & Process Development Beyond production-scale workflows, ITS AF is widely used in academic and industry research for serum reduction and serum replacement studies, primary cell culture (fibroblasts, keratinocytes, hepatocytes), hybridoma culture for antibody generation, and serum-free media development. The 10 mL and 100 mL research formats support small-scale evaluation and optimization work, with bulk and custom packaging options available when process development teams scale into manufacturing programs.

ITS Animal-Free Frequently Asked Questions (FAQs)

Product Overview

What Is ITS Animal-Free and How Does It Replace Serum in Cell Culture?

ITS Animal-Free™ (cat. no. 777ITS091) is a chemically defined, 100x liquid supplement containing recombinant insulin (1.0 g/L), recombinant human transferrin (0.55 g/L), and selenium as sodium selenite (0.67 mg/L). It replaces the core serum functions of nutrient uptake, iron delivery, and antioxidant protection in serum-free and reduced-serum media, allowing process development teams to reduce or eliminate fetal bovine serum without sacrificing cell proliferation or productivity.

How Do Insulin, Transferrin, and Selenium Each Support Cell Culture?

Recombinant insulin supports glucose uptake and metabolic signaling, recombinant transferrin delivers iron to cells through receptor-mediated endocytosis via the transferrin receptor-1 (TFR1) pathway, and selenium supports the glutathione peroxidase system that protects cells from oxidative stress. Together, these three components provide the essential non-serum inputs that most mammalian cell types require for proliferation and metabolic stability in serum-free media.

What Makes ITS Animal-Free Different From Other ITS Supplements on the Market?

ITS Animal-Free is formulated with recombinant human transferrin produced on InVitria's ExpressTec rice grain–based expression platform — the same platform behind InVitria's Optiferrin® product line and the recombinant transferrin used across major global life-science supply chains. Every component is fully recombinant or defined, with no carrier proteins, stabilizers, preservatives, or animal-derived inputs, and the product is released under the quality system of an ISO 9001:2015-certified, cGMP-compliant facility. ITS AF has also been used in clinical-stage cell therapy manufacturing, distinguishing it from research-only ITS supplements.

Clinical Use and Cell Therapy

Has ITS Animal-Free Been Used in Clinical-Stage Cell Therapy Manufacturing?

Yes. ITS Animal-Free (cat. no. 777ITS091) was used in the corneal endothelial differentiation medium for CLS001, a clinical-grade iPSC-derived cell therapy product, in a first-in-human clinical study of bullous keratopathy conducted under Japan's Act on Ensuring the Safety of Regenerative Medicine (Hirayama et al., 2025, Cell Reports Medicine). This establishes ITS AF as a supplement with demonstrated use in regulated, human-grade cell therapy manufacturing — not just research-scale applications.

Is ITS Animal-Free Suitable for CAR-T and T Cell Therapy Manufacturing?

ITS Animal-Free is compatible with T cell expansion workflows, including CAR-T manufacturing, and has been used in primary T and NK cell protocols in serum-free and reduced-serum media. For T cell therapy programs specifically requiring IL-2 supplementation alongside ITS, InVitria's OptiLeukin 2™ provides animal-origin-free recombinant human IL-2 formulated for cell therapy manufacturing.

Can ITS Animal-Free Be Used in Gene Therapy and Viral Vector Manufacturing?

Yes. ITS Animal-Free is used as a supplement in serum-free media for AAV and lentivirus production workflows in HEK293 and other production cell lines. The fully animal-free, chemically defined composition supports viral vector manufacturing programs where raw material traceability, adventitious-agent risk mitigation, and ICH Q5A compliance are priorities.

Quality and Regulatory

Is ITS Animal-Free Truly Animal-Origin-Free?

Yes. ITS Animal-Free is formulated entirely from recombinant proteins and defined inorganic components, with no animal-derived or blood-derived ingredients. Raw materials, processing aids, manufacturing equipment, and packaging are all animal-free, supporting traceable raw material documentation for regulated biomanufacturing workflows and simplifying TSE/BSE qualification in regulatory submissions.

Is ITS Animal-Free cGMP-Manufactured and ISO 9001 Certified?

Yes. InVitria manufactures ITS Animal-Free under a cGMP-compliant Quality Management System (QMS) with respect to 21 CFR Part 210/211 at its ISO 9001:2015-certified facility in Junction City, Kansas, USA. The facility and QMS support raw material qualification for IND, BLA, and supplier qualification submissions.

What Quality Documentation Is Available for ITS Animal-Free?

ITS Animal-Free is supported by a Certificate of Analysis (CoA), Safety Data Sheet (SDS), Guidelines for Use, Certificate of Origin and Animal-Free / TSE-BSE Statement, ISO 9001 certificate, and cGMP Statement. Additional quality and regulatory documentation is available on request to support specific regulatory filings.

Is ITS Animal-Free Sterile and Ready to Use?

ITS Animal-Free is supplied as a sterile-filtered 100x liquid concentrate intended for direct dilution into basal media. Re-filtration with a 0.2 µm filter is recommended if aseptic handling cannot be assured after opening or dilution.

Does ITS Animal-Free Support ICH Q5A Compliance?

The fully animal-free, chemically defined composition of ITS Animal-Free supports ICH Q5A qualification requirements for cell substrates used in the manufacture of biotechnological and biological products. The absence of animal-derived raw materials, processing aids, and packaging simplifies TSE/BSE risk assessment and reduces the documentation burden associated with raw material qualification in regulatory submissions.

Applications and Use

Does ITS Animal-Free Support Stem Cell Expansion (iPSC, HSC, MSC, CD34+)?

Yes. ITS Animal-Free has been validated in iPSC expansion with maintained pluripotency marker expression, and is used in serum-free media formulations for MSC, HSC, and CD34+ hematopoietic stem cell culture (see ITS AF iPSC expansion application note). ITS AF has also been used in clinical-stage iPSC differentiation for cell therapy manufacturing (Hirayama et al., 2025).

What Cell Types and Applications Is ITS Animal-Free Used For?

ITS Animal-Free is designed for broad mammalian cell culture use, including CHO, HEK293, VERO, hybridoma, fibroblast, keratinocyte, primary cell (T cells, NK cells, hepatocytes), and stem cell (iPSC, MSC, HSC, CD34+) workflows. It is commonly used across biologics production, cell and gene therapy manufacturing, vaccine production, and serum-free media development.

Is Recombinant ITS Better Than Plasma-Derived ITS Supplements?

Plasma-derived ITS products use transferrin isolated from human blood, which introduces donor variability, adventitious-agent risk, and additional regulatory documentation burden for TSE/BSE qualification. ITS Animal-Free replaces plasma-derived transferrin with a fully recombinant component produced on InVitria's ExpressTec platform, improving lot-to-lot consistency, eliminating blood-borne pathogen risk, and simplifying raw material qualification for regulated manufacturing.

What Concentration of ITS Animal-Free Should I Use in Serum-Free Media?

ITS Animal-Free is used at a 1x working concentration (1:100 dilution of the 100x concentrate) as a starting point. For demanding cell types, 2–3x supplementation may be appropriate. Optimal concentration depends on cell type, density, and basal medium composition — please reach out to InVitria's applications team for cell-type-specific guidance.

Selection and Comparison

What Is the Difference Between ITS, ITSE, and ITSE+A?

ITS provides insulin, transferrin, and selenium to replace core serum functions in general mammalian cell culture. ITSE adds ethanolamine to support lipid metabolism and membrane synthesis in cell types sensitive to ethanolamine limitation. ITSE+A further adds recombinant human albumin to provide broader serum-function replacement for iPSC, MSC, and other applications where additional protein stabilization is required. Select ITS AF as the baseline 3-component option, ITSE Animal-Free when ethanolamine is required, and ITSE+A for protein-stabilization-dependent workflows.

How Does ITS Animal-Free Compare to ITS-G and ITS-X Supplements on the Market?

ITS-G-style supplements (insulin, transferrin, selenium) are the classical three-component formulations, while ITS-X-style supplements add ethanolamine as a fourth component. InVitria's ITS Animal-Free aligns with the ITS-G three-component composition, while ITSE Animal-Free aligns with the ITS-X four-component composition with added ethanolamine. ITS AF is differentiated by its fully recombinant human transferrin sourced from InVitria's ExpressTec manufacturing platform and its validated use in clinical-stage cell therapy manufacturing.

Should I Use ITS Animal-Free or Individual Insulin, Transferrin, and Selenium Components?

ITS Animal-Free provides pre-formulated, pre-validated supplementation in a single sterile concentrate, reducing the media preparation steps and the number of raw materials that require independent qualification in regulated workflows. For programs that require flexibility in individual component concentrations, InVitria supplies recombinant human transferrin separately as Optiferrin®, which can be combined with recombinant insulin and selenium to formulate custom ITS preparations.

Citation and Publication

How Do I Cite ITS Animal-Free in My Methods Section?

ITS Animal-Free should be cited as: "ITS Animal-Free™ (InVitria, Junction City, KS, USA; cat. no. 777ITS091)." For publication-specific citation formatting or RRID reference support, contact InVitria's applications team.

Packaging and Ordering

What Packaging Options Are Available for ITS Animal-Free?

ITS Animal-Free is available in 10 mL, 100 mL, and 10 mL × 10 packaging formats for research and small-scale process development use. Contact InVitria for bulk and custom packaging options to support scale-up, pilot, and commercial-scale manufacturing programs.

How Can I Order ITS Animal-Free?

ITS Animal-Free is available worldwide through multiple purchasing pathways. Most customers work directly with InVitria for full access to the ITS Animal-Free product portfolio, bulk and commercial-scale supply, favorable volume-tier pricing, faster supply chain turnaround, GMP supply commitments, and regulatory documentation support. ITS Animal-Free is also available through Fisher Scientific, VWR, and the Labviva procurement platform (which integrates with Ariba, Coupa, Oracle, and Jaggaer) for customers whose institutional procurement systems require integrated supplier ordering. European customers can order with direct fulfillment from InVitria's Rotterdam, Netherlands facility. All customers receive the same product, Certificate of Analysis, and technical support regardless of channel. Contact InVitria or request bulk pricing to place an order.

Storage and Handling

What Are the Storage Conditions and Shelf Life for ITS Animal-Free?

Store ITS Animal-Free at 2–8 °C, protected from light, with containers tightly sealed. Avoid freezing. Shelf life is 2 years from date of manufacture.