TY - JOUR
T1 - Detergent-free decellularization of the human pancreas for soluble extracellular matrix (Ecm) production
AU - Tamburrini, Riccardo
AU - Chaimov, Deborah
AU - Asthana, Amish
AU - Enck, Kevin
AU - Muir, Sean M.
AU - Aziz, Justine Mariam
AU - Lablanche, Sandrine
AU - Tubbs, Emily
AU - Tomei, Alice A.
AU - Dyke, Mark Van
AU - Soker, Shay
AU - Opara, Emmanuel C.
AU - Orlando, Giuseppe
N1 - Funding Information:
This project has received funding from the European Union?s Horizon 2020 Research and Innovation Program under grant agreement No. 646272. Human Pancreatic Islets were obtained from Prodo Laboratories, Aliso Viejo, CA 92656.
Funding Information:
This project has received funding from the European Union’s Horizon 2020 Research and Innovation Program under grant agreement No. 646272. Human Pancreatic Islets were obtained from Prodo Laboratories, Aliso Viejo, CA 92656.
Publisher Copyright:
© 2020 JoVE Journal of Visualized Experiments.
PY - 2020/9
Y1 - 2020/9
N2 - Islet transplantation (ITx) has the potential to become the standard of care in beta cell replacement medicine but its results remain inferior to those obtained with whole pancreas transplantation. The protocols currently used for human islet isolation are under scrutiny because they are based on the enzymatic digestion of the organ, whereby the pancreas is demolished, its connections to the body are lost and islets are irreversibly damaged. Islet damage is characterized by critical factors such as the destruction of the extracellular matrix (ECM), which represents the 3D framework of the islet niche and whose loss is incompatible with islet euphysiology. Researchers are proposing the use of ECM-based scaffolds derived from the mammalian pancreas to address this problem and ultimately improve islet viability, function, and lifespan. Currently available methods to obtain such scaffolds are harsh because they are largely detergent based. Thus, we propose a new, detergent-free method that creates less ECM damage and can preserve critical components of pancreatic ECM. The results show that the newly developed decellularization protocol allowed the achievement of complete DNA clearance while the ECM components were retained. The ECM obtained was tested for cytotoxicity and encapsulated with human pancreatic islets which showed a positive cellular behavior with insulin secretion when stimulated with glucose challenge. Collectively, we propose a new method for the decellularization of the human pancreas without the use of conventional ionic and non-ionic chemical detergents. This protocol and the ECM obtained with it could be of use for both in vitro and in vivo applications.
AB - Islet transplantation (ITx) has the potential to become the standard of care in beta cell replacement medicine but its results remain inferior to those obtained with whole pancreas transplantation. The protocols currently used for human islet isolation are under scrutiny because they are based on the enzymatic digestion of the organ, whereby the pancreas is demolished, its connections to the body are lost and islets are irreversibly damaged. Islet damage is characterized by critical factors such as the destruction of the extracellular matrix (ECM), which represents the 3D framework of the islet niche and whose loss is incompatible with islet euphysiology. Researchers are proposing the use of ECM-based scaffolds derived from the mammalian pancreas to address this problem and ultimately improve islet viability, function, and lifespan. Currently available methods to obtain such scaffolds are harsh because they are largely detergent based. Thus, we propose a new, detergent-free method that creates less ECM damage and can preserve critical components of pancreatic ECM. The results show that the newly developed decellularization protocol allowed the achievement of complete DNA clearance while the ECM components were retained. The ECM obtained was tested for cytotoxicity and encapsulated with human pancreatic islets which showed a positive cellular behavior with insulin secretion when stimulated with glucose challenge. Collectively, we propose a new method for the decellularization of the human pancreas without the use of conventional ionic and non-ionic chemical detergents. This protocol and the ECM obtained with it could be of use for both in vitro and in vivo applications.
UR - http://www.scopus.com/inward/record.url?scp=85090663842&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85090663842&partnerID=8YFLogxK
U2 - 10.3791/61663
DO - 10.3791/61663
M3 - Article
C2 - 32955501
AN - SCOPUS:85090663842
VL - 2020
SP - 1
EP - 15
JO - Journal of visualized experiments : JoVE
JF - Journal of visualized experiments : JoVE
SN - 1940-087X
IS - 163
M1 - e61663
ER -