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Spider silk exhibits exceptional strength and elasticity in its natural form. Over the course of several decades, researchers have been working on artificially spinning recombinant spider silk proteins (spidroin) in order to replicate the remarkable mechanical properties of natural spider silk. In this study, we utilized the wet spinning method to investigate the relationship between the concentration of the coagulation bath and fiber performance. We discovered that the concentration of methanol plays a crucial role in determining the continuity, diameter, and mechanical properties of the fibers. Lower concentrations of methanol favor the production of continuous, thinner fibers with higher strain. Additionally, secondary stretching during the spinning process contributes to the production of silk fibers with stable mechanical properties and thermal stability. By employing different concentrations of methanol and applying additional stretching, we successfully produced silk fibers with a high strain of 2.1652 ± 0.3871 mm/mm. Furthermore, these wet-spun fibers demonstrated the ability to promote the growth of Schwann cells, indicating their potential application in the field of biomedical engineering. Hence, the exceptional mechanical properties and the ability to promote cellular growth make the obtained spider silk fibers highly promising for various biomedical applications. [ABSTRACT FROM AUTHOR]