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Small intestinal submucosa: a substrate for in vitro cell growth
S F Badylak1, R Record, K Lindberg
1Hillenbrand Biomedical Engineering Center, Purdue University, West Lafayette, IN 47907, USA. badylak@ecn.purdue.edu
Abstract:
The extracellular matrix (ECM) of the small intestinal submucosa (SIS) was harvested by removing the superficial layers of the mucosa and the external muscular layers. The remaining 80 microns thick sheet was disinfected and sterilized by methods which removed all cellular components. The SIS-ECM, retaining its native 3-dimensional microarchitecture and composition, was evaluated for its ability to support in vitro cell growth. Six separate cell types were seeded either alone or in coculture with other cells upon this matrix, grown in selected media, a examined daily for time periods ranging from 48 h to 2 weeks. The six cell types tested were NIH Swiss mouse 3T3 fibroblast, NIH 3T3/j2 fibroblasts, primary human fibroblasts, primary human keratinocytes, human microvascular endothelial cells (HMECs), and an established rat osteosarcoma (ROS) cell line. All cell types showed the ability to attach a proliferate. All fibroblast cell line and the keratinocytes proliferated and/or migrated into the 3-dimensional scaffold of the SIS matrix. The ROS cells and the HMECs were confined in their growth pattern to the surface of the matrix. Coculturing of NIH 3T3/J2 fibroblasts and primary human keratinocytes resulted in a distinctive spatial orientation of the two cell types. The fibroblast populated the mid-substance of the 3-dimensional matrix and the keratinocytes formed an epidermal structure with rete ridge-like formation and stratification when the composite was lifted to an air liquid interface in culture. In summary, SIS provides a substratum with a 3-dimensional scaffold that allows for cell migration and spatial organization. The substratum is suitable for in vitro studies of the interaction between epithelial or mesenchymal cells and a naturally occurring extracellular matrix.
Insights
Small intestinal submucosa extracellular matrix (SIS-ECM) supports diverse cell growth and organization in vitro. This 3D scaffold is ideal for studying cell interactions with natural matrices.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Cell Biology
Background:
- The extracellular matrix (ECM) plays a crucial role in cell behavior and tissue development.
- Small intestinal submucosa (SIS) is a natural ECM with potential applications in regenerative medicine.
Purpose of the Study:
- To evaluate the ability of SIS-derived ECM (SIS-ECM) to support in vitro growth and organization of various cell types.
- To investigate the potential of SIS-ECM as a 3D scaffold for cell-based studies.
Main Methods:
- SIS-ECM was prepared by removing mucosal and muscular layers, followed by disinfection and sterilization.
- Six cell types (fibroblasts, keratinocytes, endothelial cells, osteosarcoma cells) were cultured on SIS-ECM alone or in co-culture.
- Cell attachment, proliferation, migration, and spatial organization were assessed over 48 hours to 2 weeks.
Main Results:
- All tested cell types attached and proliferated on the SIS-ECM.
- Fibroblasts and keratinocytes migrated into the 3D SIS-ECM scaffold.
- Osteosarcoma cells and microvascular endothelial cells showed surface-confined growth patterns.
- Co-culture of fibroblasts and keratinocytes resulted in distinct spatial organization and epidermal structure formation.
Conclusions:
- SIS-ECM provides a 3D scaffold that supports cell migration and spatial organization.
- SIS-ECM is a suitable substratum for in vitro studies of cell-matrix interactions.
- The native microarchitecture and composition of SIS-ECM facilitate complex cellular behaviors.