Organ-specific ECM arrays for investigating cell-ECM interactions during stem cell differentiation

Abstract

Pluripotent stem cells are promising source of cells for tissue engineering, regenerative medicine and drug discovery applications. The process of stem cell differentiation is regulated by multi-parametric cues from the surrounding microenvironment, one of the critical one being cell interaction with extracellular matrix (ECM). The ECM is a complex tissue-specific structure which is an important physiological regulator of stem cell function and fate. Recapitulating this native ECM microenvironment niche is best facilitated by decellularized tissue/organ derived ECM, which can faithfully reproduce the physiological environment with high fidelity to in vivo condition and promote tissue-specific cellular development and maturation. Recognizing the need for organ specific ECM in a 3D culture environment in driving phenotypic differentiation and maturation of hPSCs, we fabricated an ECM array platform using native-mimicry ECM from decellularized organs (namely pancreas, liver and heart), which allows cell-ECM interactions in both 2D and 3D configuration. The ECM array was integrated with rapid quantitative imaging for a systematic investigation of matrix protein profiles and sensitive measurement of cell-ECM interaction during hPSC differentiation. We tested our platform by elucidating the role of the three different organ-specific ECM in supporting induced pancreatic differentiation of hPSCs. While the focus of this report is on pancreatic differentiation, the developed platform is versatile to be applied to characterize any lineage specific differentiation.

Mimicking Physiologically Relevant Hepatocyte Zonation Using Immunomodulatory Silk Liver Extracellular Matrix Scaffolds toward a Bioartificial Liver Platform

“…Liver-ECM-based 3D matrices are a promising platform for regulating physiological attributes of cultured hepatocytes, which reproduce native ECM microenvironment niche through multiparametric cues, thus promoting cellular maturation and cell–ECM interaction . However, individual ECM protein has proven to be inadequate in maintaining a native cell phenotype and promoting lineage-specific differentiation of progenitor cells compared to tissue-specific ECM. ,, To facilitate hepatocyte functions, several groups have fabricated liver-ECM-based 2D and 3D platforms.…”

Preparation and Use of Decellularized Extracellular Matrix for Tissue Engineering

“…For many solid organs and tissues, canulating and perfusing the primary artery is one of the best approaches. The kidney is perfused through the renal artery [ 148 , 164 , 243 ], the liver through the portal vein and/or the hepatic artery [ 169 , 245 ], the pancreas through the splenic vein [ 99 ], the uterus through the uterine artery [ 153 ], and the corpus cavernosum through the cavernosal artery [ 157 ]. Other organs that are typical decellularized through chemical baths can be decellularized if there is a need to keep the structure intact, such as perfusing the superior mesenteric artery to decellularize the jejunum [ 144 ], the common carotid artery for the cervical esophagus [ 246 ], and the ureter for bladders [ 185 ].…”

“…Stem cells have the unique ability to differentiate into different tissue types, but they generally require specialized treatments to induce this differentiation into a particular cell type. The specific make-up of the ECM is unique to each tissue type, and the ECM has been shown to have the potential to induce and enhance the differentiation of stem cells towards the phenotype associated with the tissue source of the ECM [ 14 , 56 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 100 , 101 , 102 , 103 , 104 , 105 ].…”

Repopulation of decellularized organ scaffolds with human pluripotent stem cell-derived pancreatic progenitor cells

“…In contrast, Citro et al by demonstrated the feasibility of reengineering the lung scaffold to house pancreatic islets [61], which opens up a new paradigm in engineered tissue. In our previous work we have demonstrated compatibility of non-pancreatic ECM with hPSC-PP cells [62]. This prompted us to consider alternate tissue scaffolds to re-engineer the pancreas.…”

“…In addition, liver originates from endodermal germ layer with close anatomical proximity and association with pancreas. Further, our recent work demonstrated the compatibility of liver-ECM for supporting pancreatic differentiation [62]. To this end, we hypothesized that decellularized liver could yield an alternate compatible organ scaffold to support the repopulation of hPSC PP-aggregates.…”