Serum is an indispensable factor in the maintenance of the biological characteristics of sweat gland cells.

The tolerance of sweat gland cells for in vitro amplification and subcultivation is low as they are somatic cells. The present study aimed to formulate an optimal medium for the culture of human eccrine sweat gland cells (HESGCs) and to establish a method for induction of HESGCs proliferation, whilst maintaining the characteristics of sweat gland cells. HESGCs cultured in sweat gland (SG):keratinocyte growth medium‑2 (KGM‑2) (1:1) medium had a higher proliferation rate and a stable morphology compared with cells cultured in SG and KGM‑2 medium only. Reverse transcription‑quantitative polymerase chain reaction indicated that cells cultured in the SG:KGM‑2 (1:1) medium exhibited higher expression levels of α‑smooth muscle actin, keratin (K)77, carcinoembryonic antigen, K8, K18, ectodysplasin A receptor, c‑Myc, Kruppel‑like factor 4 and octamer‑binding transcription factor 4 compared with cells cultured in SG only or KGM‑2 only medium. Three‑dimensional culture analysis revealed that HESGCs cultured in SG:KGM‑2 1:1 medium differentiated into sweat gland‑like structures, whereas cells cultured in KGM‑2 only medium underwent cornification. The present study also determined that the maintenance of the biological characteristics of HESGCs occurred due to the presence of fetal bovine serum (FBS). Cells cultured in medium without FBS differentiated into keratinocytes. Therefore, the SG:KGM‑2 (1:1) medium may be a suitable culture medium for HESGCs. In conclusion, this mixed medium is a valuable compound and should be considered to be a potential supplemental medium for HESGCs.