Stephanie Louise Kwolek was born in New Kensington, Pennsylvania, to Polish immigrant parents John Kwolek and Nellie (Zajdel) Kwolek. Her father, a naturalist, instilled in her a love for the natural world, while her mother, a seamstress, nurtured her creativity and appreciation for design. These early influences shaped her inquisitive mind and attention to detail, qualities that would prove essential to her future success as a chemist.

Stephanie Kwolek graduated from Margaret Morrison Carnegie College, now part of Carnegie Mellon University, with a Bachelor of Science degree in Chemistry. She originally intended to become a doctor and hoped to earn enough money to attend medical school. Her strong academic performance and passion for chemistry made her a highly sought-after candidate for various research positions.