Differentiation of Menstrual Blood Stem Cells into Keratinocyte-Like Cells on Bilayer Nanofibrous Scaffold

Abstract

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.

Endometrial and Menstrual Blood Mesenchymal Stem/Stromal Cells: Biological Properties and Clinical Application

“…MenSC seeded on an amniotic membrane improved wound closure in a rat excisional wound defect model (Farzamfar et al, 2018). In vitro, MenSCs differentiated into keratinocyte-like cells (Akhavan-Tavakoli et al, 2017), an effect potentiated in 3D culture (Fard et al, 2018) or in a biomimetic nanofibrous scaffold, two approaches that increased the wound healing capacity of MenSC (Arasteh et al, 2018). Such data imply the importance of ECM in the differentiation of MenSC into keratinocyte-like cells and further highlight the value of MenSCs as a potential therapeutic in clinical settings.…”

“…The MenSC-derived keratinocytes expressed keratinocytespecific markers K14, p63 and IVL (involucrin). Generating keratinocytes from MenSC on an efficient natural construct has potential applicability for MSC-based skin wound healing and regeneration (Arasteh et al, 2018;Fard et al, 2018), although these tissue engineering constructs have yet to be evaluated in an animal skin wound repair model. MenSC show considerable capacity to differentiate into numerous lineages in vitro, although have lesser ability for several bmMSC lineages.…”

“…Unfractionated MenSC have potential to differentiate into keratinocyte-like cells, generating epidermal lineage markers via co-culturing with keratinocytes derived from the foreskin of healthy newborns ( Faramarzi et al, 2016 ; Akhavan-Tavakoli et al, 2017 ). MenSC were also induced to differentiate into keratinocyte-like cells in 3D culture with human foreskin-derived keratinocytes on a bilayer scaffold composed of amniotic membrane and silk fibroin ( Arasteh et al, 2018 ; Fard et al, 2018 ). The MenSC-derived keratinocytes expressed keratinocyte-specific markers K14 , p63 and IVL (involucrin).…”

Non-gynaecological Applications of Menstrual-derived Stem Cells: A Systematic Review

“…MenSC can differentiate into several lineages including endothelial, osteogenic, chondrogenic, adipocytic, pancreatic, hepatic, respiratory epithelial, cardiomyocytic, and neurocytic lineages 1 , 7 – 10 . MenSC have shown potential therapeutic application in various non-gynaecological disorders including wound healing 11 – 14 , neurological 15 – 17 , musculoskeletal 18 , 19 , cardiovascular 5 , 20 , 21 , respiratory 5 , 20 , 22 and liver 23 , 24 diseases, as well as in diabetes 25 , 26 and cancer 27 – 31 . Several studies also considered the gynaecological applications of MenSC, such as in ovarian-related diseases 32 , 33 , endometrial injury 34 , 35 and Asherman syndrome 36 , 37 .…”

“…The exclusion criteria used in various studies were: vaginal infection history, non-steroidal anti-inflammatory drugs, corticosteroids and oral contraceptives use within the last 3 months, infections such as human immunodeficiency viruses, hepatitis C virus and hepatitis B virus, autoimmune diseases, diabetes, endometriosis and malignancies 63 – 66 . Menstrual blood for the isolation of MenSC should be collected on the day with the heaviest blood flow 65 – 66 usually the second day 64 , 67 – 69 . Around 5 ml volume of blood will provide sufficient stem cells.…”