108) This method can be used to identify optimal decellularization protocols for recellularization of three-dimensional tissue-scaffolds with embryonic stem cells and other tissue-specific cell types. |
PMID:23172824 DOI:10.1002/term.1658 |
2015 Journal of tissue engineering and regenerative medicine |
* Dewaxed ECM: A simple method for analyzing cell behaviour on decellularized extracellular matrices. |
- Decellularization techniques have been used on a wide variety of tissues to create cell-seedable scaffolds for tissue engineering. Finding a suitable decellularization protocol for a certain type of tissue can be laborious, especially when organ perfusion devices are needed. In this study, we report a quick and simple method for comparing decellularization protocols combining the use of paraffin slices and two-dimensional cell cultures. We developed three decellularization protocols for adult murine kidney that yielded decellularized extracellular matrices (ECMs) with varying histological properties. The resulting paraffin-embedded ECM slices were deparaffinized and reseeded with murine embryonic stem cells (mESCs). We analyzed cell attachment four days post seeding via determination of cell numbers, and used quantitative Real-Time PCR 13 days post seeding to measure gene expression levels of two genes associated with renal development, Pax2 and Pou3f3. The three decellularization protocols produced kidney-matrices that showed clearly distinguishable results. We demonstrated that formerly paraffin-embedded decellularized ECMs can effectively influence differentiation of stem cells. This method can be used to identify optimal decellularization protocols for recellularization of three-dimensional tissue-scaffolds with embryonic stem cells and other tissue-specific cell types. |
(1)80 *null* | (15)5 as | (29)3 exposed | (43)2 expanded |
(2)47 and | (16)5 expressed | (30)3 have | (44)2 expressing |
(3)39 were | (17)5 of | (31)3 isolated | (45)2 had |
(4)26 in | (18)5 which | (32)3 showed | (46)2 labeled |
(5)12 (MSCs) | (19)4 (ASCs) | (33)3 we | (47)2 may |
(6)11 was | (20)4 has | (34)2 (ASCs), | (48)2 play |
(7)10 to | (21)4 is | (35)2 (BMSCs) | (49)2 present |
(8)9 with | (22)4 on | (36)2 (ECFCs) | (50)2 remains |
(9)7 are | (23)4 seeded | (37)2 (HSCs) | (51)2 such |
(10)6 can | (24)4 within | (38)2 after | (52)2 the |
(11)6 for | (25)3 (ASC) | (39)2 by | (53)2 via |
(12)6 from | (26)3 (ECs) | (40)2 caused | |
(13)6 or | (27)3 (HUVECs) | (41)2 cultured | |
(14)6 that | (28)3 (hMSCs) | (42)2 derived |
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