Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
Biomedical Engineering Department, University of Connecticut, Hartford, CT, USA.
Methods Mol Biol. 2020;2125:129-156. doi: 10.1007/7651_2018_193.
Skin tissue engineering is a high-throughput technology to heal the wounds. Already, considerable advances have been achieved using stem cells for wound healing applications. Menstrual blood stem cell (MenSC) is an available and accessible source of stem cells that have differentiation potential into a wide range of lineages like keratinocytes. Extracellular matrix like substratum plays an impressive role in skin regeneration as an attachment site for stem cells by transmitting the bioactive signals and provoking stem cells to differentiate into keratinocyte lineage. The biomimetic nanofibrous scaffold especially in bilayer format has been extensively utilized to develop skin equivalents. This chapter explains detailed protocols of keratinocyte differentiation of MenSCs on bilayer scaffold comprising amniotic membrane and fibroin nanofibers. The isolated MenSCs are seeded on the nanofibers and subsequently differentiated into keratinocyte lineage in co-culture with foreskin-derived keratinocytes. Immunofluorescence staining is used to evaluate the development of seeded MenSCs in bilayer scaffold into keratinocyte-like cells.
皮肤组织工程是一种高通量技术，可用于治疗伤口。目前，已经在使用干细胞治疗伤口应用方面取得了相当大的进展。月经血干细胞（MenSC）是一种可用且可获得的干细胞来源，具有向广泛谱系分化的潜力，如角质细胞。细胞外基质如基底膜在皮肤再生中起着重要作用，作为干细胞的附着点，通过传递生物活性信号并促使干细胞分化为角质细胞谱系。仿生纳米纤维支架，特别是双层形式，已被广泛用于开发皮肤等效物。本章详细介绍了在包含羊膜和丝素纳米纤维的双层支架上 MenSC 向角质细胞分化的方案。分离的 MenSC 接种在纳米纤维上，然后与包皮来源的角质细胞共培养，分化为角质细胞谱系。免疫荧光染色用于评估接种在双层支架上的 MenSC 向角质细胞样细胞的发育。
