354) 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. |
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)101 and | (12)8 including | (23)4 for | (34)2 13 |
(2)36 *null* | (13)7 encoding | (24)4 from | (35)2 but |
(3)28 are | (14)7 involved | (25)4 related | (36)2 can |
(4)26 were | (15)7 which | (26)4 with | (37)2 coding |
(5)25 in | (16)6 (PCGs), | (27)3 had | (38)2 expressed |
(6)16 2 | (17)6 use | (28)3 have | (39)2 is |
(7)14 (13 | (18)5 may | (29)3 stop | (40)2 on |
(8)14 22 | (19)5 that | (30)3 such | (41)2 ranged |
(9)12 start | (20)4 (PCGs) | (31)2 (cytb, | (42)2 to |
(10)11 of | (21)4 as | (32)2 (pnad1 | |
(11)9 was | (22)4 associated | (33)2 (srfAA, |
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