pmid: 31721538,   nlmid 101566618,  doi:10.22074/cellj.2020.6604

Optimizing The Cell Seeding Protocol to Human Decellularized Ovarian Scaffold: Application of Dynamic System for Bio-Engineering.

Mirzaeian, Leila Eivazkhani, Farideh Hezavehei, Maryam Moini, Ashraf Esfandiari, Fereshteh Valojerdi, Mojtaba Rezazadeh Fathi, Rouhollah

Vol. 22, Issue 22, Jul.2020

Cell journal (Cell J),  ISSN 2228-5806


Decellularized tissue scaffolds provide an extracellular matrix to control stem cells differentiation toward specific lineages. The application of mesenchymal stem cells for artificial ovary production may enhance ex vivo functions of the ovary. On the other hand, the scaffold needs interaction and integration with cells. Thus, the development of ovarian engineered constructs (OVECs) requires the use of efficient methods for seeding of the cells into the ovarian and other types of scaffolds. The main goal of the present study was to develop an optimized culture system for efficient seeding of peritoneum mesenchymal stem cells (PMSCs) into human decellularized ovarian scaffold. In this experimental study, three methods were used for cellular seeding including rotational (spinner flask) and static (conventional and injection) seeding cultures. OVECs were evaluated with Hematoxylin and Eosin staining and viability analyses for the seeded PMSCs. Then, immunohistochemistry analysis was performed using the best method of cellular seeding for primordial germ cell-like cells, mesenchymal stem cells and proliferation markers. Stereology analysis was also performed for the number of penetrated cells into the OVECs. Our results showed that rotational seeding increases the permeability of PMSCs into the scaffold and survival rate of the seeded PMSCs, comparing to the other methods. On the other hand, rotationally seeded PMSCs had a more favorable capability of proliferation with Ki67 expression and differentiation to ovarian specific cells with expression of primordial germ cell line markers without mesenchymal stem cells markers production. Furthermore, stereology showed a more favorable distribution of PMSCs along the outer surfaces of the OVEC with further distribution at the central part of the scaffold. The average total cell values were determined 2142187 cells/mm on each OVEC. The rotational seeding method is a more favorable approach to cell seeding into ovarian decellularized tissue than static seeding.

MESH Headings

Article Keywords

Mesenchymal Stem Cells | Ovary | Peritoneum | Seeding | Tissue Engineering |


Similar Articles

In vitro cartilage construct generation from silk fibroin- chitosan porous scaffold and umbilical cord blood derived human mesenchymal stem cells in dynamic culture condition.
Agrawal, Parinita Pramanik, Krishna Biswas, Amit Ku Patra, Ranjan
Journal of biomedical materials research. Part A ,vol(106), issue(2),  Feb.2018

Mammalian neo-oogenesis and expression of meiosis-specific protein SCP3 in adult human and monkey ovaries.
Bukovsky, Antonin Caudle, Michael R Gupta, Satish K Svetlikova, Marta Selleck-White, Rachel Ayala, Alena M Dominguez, Roberto
Cell cycle (Georgetown, Tex.) ,vol(7), issue(5),  Mar.2008

[Experimental study on isolation and cultivation of placenta-derived mesenchymal stem cells and bone marrow-derived mesenchymal stem cells of rabbit and their biological characteristics].
Lu, Yao Deng, Li Li, Xiuqun
Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery ,vol(21), issue(9),  Sep.2007

Novel Action of FSH on Stem Cells in Adult Mammalian Ovary Induces Postnatal Oogenesis and Primordial Follicle Assembly.
Bhartiya, Deepa Parte, Seema Patel, Hiren Sriraman, Kalpana Zaveri, Kusum Hinduja, Indira
Stem cells international ,vol(2016), issue(),  

Osteoclastogenesis on tissue-engineered bone.
Nakagawa, Keisuke Abukawa, Harutsugi Shin, Michael Y Terai, Hidetomi Troulis, Maria J Vacanti, Joseph P
Tissue engineering ,vol(10), issue(1-2),  

Differentiation within autologous fibrin scaffolds of porcine dermal cells with the mesenchymal stem cell phenotype.
de la Puente, Pilar Ludeña, Dolores López, Marta Ramos, Jennifer Iglesias, Javier
Experimental cell research ,vol(319), issue(3),  Feb.2013

Gonadotropin treatment augments postnatal oogenesis and primordial follicle assembly in adult mouse ovaries?
Bhartiya, Deepa Sriraman, Kalpana Gunjal, Pranesh Modak, Harshada
Journal of ovarian research ,vol(5), issue(1),  Nov.2012

Self-assembled composite matrix in a hierarchical 3-D scaffold for bone tissue engineering.
Chen, Muwan Le, Dang Q S Baatrup, Anette Nygaard, Jens V Hein, San Bjerre, Lea Kassem, Moustapha Zou, Xuenong Bünger, Cody
Acta biomaterialia ,vol(7), issue(5),  May.2011

A biaxial rotating bioreactor for the culture of fetal mesenchymal stem cells for bone tissue engineering.
Zhang, Zhi-Yong Teoh, Swee Hin Chong, Woon-Shin Foo, Toon-Tien Chng, Yhee-Cheng Choolani, Mahesh Chan, Jerry
Biomaterials ,vol(30), issue(14),  May.2009

Initiation of puberty in mice following decellularized ovary transplant.
Laronda, Monica M Jakus, Adam E Whelan, Kelly A Wertheim, Jason A Shah, Ramille N Woodruff, Teresa K
Biomaterials ,vol(50), issue(),  May.2015

Medline data is available here.

The Search Enterprise

Contact Us