Rousselot, Darling Ingredients’ health brand and expert of collagen-based solutions, revealed the results of a new study which demonstrates that Rousselot’s ultra-pure GMP-grade gelatins outperformed standard non-endotoxin purified research-grade gelatins and other coating materials in 2D cell culture. Data demonstrate that endotoxin (lipopolysaccharide, LPS) levels and the nature of biomaterials used in cell coating have a significant impact on cell morphology and proliferation.
2D cell culture involves securing, nourishing, and growing cells on a flat surface. Most cells in the body are attached or anchored to an extracellular matrix (ECM) that consists of proteins like collagen, fibronectin, and laminin. Therefore, most cells require a coating with an ECM material that resembles their natural environment to improve cell attachment, proliferation, and sometimes cellular performance. A nourishing cellular environment means reducing factors that can lead to cellular stress. Multiple studies have shown that even traces of endotoxins can induce serious adverse effects on stem cell behavior and function.
To secure and grow cells, Rousselot used gelatin, a biomaterial derived from collagen, to recreate the natural cellular environment within a laboratory setting. Rousselot investigated the effect of three different coatings on the morphology and proliferation of Adipose-Derived Mesenchymal Stem Cells (AD MSCs):
- Rousselot X-Pure gelatin, characterized by low endotoxin levels (<10 EU/g) and 155 kDa molecular weight
- A commercially available standard research-grade gelatin, with endotoxin levels of 4100 EU/g and molecular weight of 160kDa
- Standard fibronectin coating material, with endotoxin levels of 20000 EU/g and molecular weight of 220 kDa
The cell morphology of cells grown on discs coated with X-Pure or commercial gelatin showed elongated and aligned cells with a spindle-shaped morphology, whereas the cells on fibronectin were less dense with more diversity in shape. In addition, Rousselot’s X-Pure gelatin-coating resulted in approximately 70% increased cell growth (measured as lactate production during a 7-day culture period) compared to fibronectin. The research-grade, non-purified gelatin-coating only resulted in an approximately 35% increase (see graph).
“The use of low endotoxin gelatin should be standard practice in cell culture applications,” commented Jos Olijve, Scientific Support Manager at Rousselot. “Not only it is essential to nurture and promote cell growth to achieve consistent and reliable data, but as you move from basic cell therapy research to the clinic, low endotoxin products with complete documentation and traceability are essential for regulatory approval. Using purified, consistently produced, and high-quality biomaterials throughout product development minimize the risk of unexpected and inconsistent results and speed up the translation.”
