Tissue Engineered Human Amniotic Membrane Application in Mouse Ovarian Follicular Culture

Abstract

Since folliculogenesis requires a powerful cell-matrix interaction, natural scaffolds seem to be needed for follicular culture. Human amniotic membrane (HAM) offers promise as a support of in vitro ovarian follicular culture. HAM was decellularized with trypsin and EDTA. DNA and histology assays were performed to determine the elimination rate of genomic components. Cyto-biocompatibility of decellular AM (DAM) was verified by the cell viability (MTT) test. The small parts of intact amniotic membrane (IAM) and DAM were coated on the bottom of 96-well and each well was filled with 150 µL of base medium. Mouse primary-secondary (PS) follicles were separated to three groups: 1-culture in base medium (Control), 2-culture on IAM and 3-culture on DAM. Follicular size, morphology, viability, estradiol production and genes expression were evaluated and IAM group showed better growth and development in follicle culture. The viability rate and estradiol production in both experimental groups were statistically higher than the Control. Gdf9, Bmp15 and Cx37 were found to have higher expression levels in IAM group. Also, maximum apoptotic and survival indexes were determined in Control and IAM groups, respectively. Finally, IAM provides a better protective environment for mouse PS follicular culture that can reduce apoptosis level.

Tissue-Engineered Grafts from Human Decellularized Extracellular Matrices: A Systematic Review and Future Perspectives

“…In particular, acellular amniotic membrane was investigated as a scaffold to support/deliver epithelial cells [240], limbal stem cells [296], human adipose stem cells [297], differentiated neural-like cells [298], mice ovarian follicular culture [299], as well as a scaffold with cell-guiding ability [300] but it may also be used as a surgical patch/mesh [301] for reconstruction of vessels [302] (also once rolled up [303,304]); bladder [305] or circumferential urethral defect [306], reconstruction of esophageal wall [307], cardiac [120,308] and cerebrospinal fluid [308] applications, endometrial fibrosis treatment [309], soft tissue damage, limbal stem cell deficiency [293], skin defects also due to ulcers or severe skin burns or for corneal defects [310,311,312], treatment of pharyngocutaneous fistula after total laryngectomy [291], and for fetoscopic closure of iatrogenic defects in fetal membranes [313].…”

“…Several decellularization methods were approached, often combining multiple phases based on physical (freezing–thawing), osmotic-chemical (Triton X-100, SDS, EDTA, Tris, NaOH, NH 4 Cl) and/or enzymatic (trypsin, dispase, lipase, DNase, and RNase) treatments [120,234,289,291,292,293,294,296,297,298,299,300,301,302,303,304,305,306,307,308,309,311,312,313,314,315,317,318,319].…”

“…Various nonstem cells were also investiganted and showed good repopulation properties: rabbit macrophages [293]; murine HL-1 cells and human immune cells derived from buffy coat [144]; mice primary follicles [299]; human keratinocytes [311]; porcine oral epithelial cells [307]; human amniotic epithelial cells [298,313]; fibroblasts (mouse embryonic—NIH3T3; human cardiac, foreskin, dermal, and nasal turbinate) [144,311,313,316,317,318]; rabbit, mouse, and human smooth muscle cells [301,305]; cardiac myocytes, [302]; human skeletal muscle cells [300]; human endothelial cells [302,303,313]; human chondrocytes [313].…”

Special Issue Devoted to a New Field of Regenerative Medicine: Reproductive Tissue Engineering

“…19 The largest area of interest to emerge in the field of REPROTEN has been fertility restoration in female patients, as indicated by the numerous papers on the subject in this special issue, mainly focusing on ovarian tissue engineering. Studies by our group 2,20 and Motamed et al 14 have demonstrated that different scaffolds, using materials like fibrin, alginate and amniotic membranes, can be successfully applied to grow isolated preantral follicles. He 9 and David et al 5 developed exciting new strategies to encapsulate isolated follicles or fragments of ovarian tissue for further in vitro culture or transplantation to restore fertility or endocrine function.…”

Co‐culture with peritoneum mesothelial stem cells supports the in vitro growth of mouse ovarian follicles

“…conditioned media are other techniques that are being investigated for better in vivo development of follicles. 6,[9][10][11] Many studies have used different synthetic (such as poly ethylene glycol 12 and polyvinyl alcohol 13 ), and natural matrices (such as agar, alginate, 14 derivation of hyaluronan, 15 and fibrin 16 ), as well as tissuederived extracellular matrix (ECM) (such as amniotic membranes 17 ) for in vitro culture of follicles. Synthetic matrices have limitations in terms of their biomechanical properties and are associated with high production costs.…”