Impaired wound healing in critical limb ischemia (CLI) results from multiple factors that affect many cell types and their behavior. Epidermal keratinocytes and dermal fibroblasts play crucial roles in wound healing. However, it remains unclear whether these cell types irreversibly convert into a non-proliferative phenotype and are involved in impaired wound healing in CLI. Here, we demonstrate that skin keratinocytes and fibroblasts isolated from CLI patients maintain their proliferative potentials. Epidermal keratinocytes and dermal fibroblasts were isolated from the surrounding skin of foot wounds in CLI patients with diabetic nephropathy on hemodialysis, and their growth potentials were evaluated. It was found that keratinocytes from lower limbs and trunk of patients can give rise to proliferative growing colonies and can be serially passaged. Fibroblasts can also form colonies with a proliferative phenotype. These results indicate that skin keratinocytes and fibroblasts maintain their proliferative capacity even in diabetic and ischemic microenvironments and can be reactivated under appropriate conditions. This study provides strong evidence that the improvement of the cellular microenvironments is a promising therapeutic approach for CLI and these cells can also be used for potential sources of skin reconstruction.

严重肢体缺血 (CLI) 中的伤口愈合受损是由影响许多细胞类型及其行为的多种因素造成的。表皮角质形成细胞和真皮成纤维细胞在伤口愈合中起着至关重要的作用。然而，目前尚不清楚这些细胞类型是否不可逆地转化为非增殖表型并参与 CLI 中伤口愈合受损。在这里，我们证明从 CLI 患者中分离的皮肤角质形成细胞和成纤维细胞保持其增殖潜力。从血液透析的糖尿病肾病 CLI 患者足部伤口周围皮肤中分离表皮角质形成细胞和真皮成纤维细胞，并评估其生长潜力。研究发现，来自患者下肢和躯干的角质形成细胞可以产生增殖性生长的集落，并且可以连续传代。成纤维细胞还可以形成具有增殖表型的集落。这些结果表明，即使在糖尿病和缺血微环境中，皮肤角质形成细胞和成纤维细胞也能保持其增殖能力，并且可以在适当的条件下重新激活。这项研究提供了强有力的证据，证明细胞微环境的改善是 CLI 的一种有前景的治疗方法，并且这些细胞也可用于皮肤重建的潜在来源。