We would like to thank Dr. George Commons and Dr. Dean Vistnes for their support and collaboration of our research. This work is supported by National Institute of Dental and Craniofacial Research grants 1 R21 DE019274-01, R01EB009689 and RC2 DE020771- 02, the Oak Foundation and Hagey Laboratory for Pediatric Regenerative Medicine to M.T.L. Dr. Hyun is supported by the Saint Joseph Mercy Hospital GME.

1. Cell Isolation &amp; Expansion
<ol>
 <li>All patient consent and experimental protocols were reviewed and approved by Stanford University Institutional Review Board (Protocol #2188 and #9999).</li>
 <li>Obtain human subcutaneous adipose tissue from elective lipoaspiration procedures under local/general anesthesia.</li>
 <li>There will be two layers in the lipoaspirate (Figure 1A). The supernatant contains the vast majority of the processed cellular material. The bottom layer is mostly injected salin...

<ol>
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Since the isolation of adipose-derived stromal cells2 from lipoaspirate, these cells have been differentiated into a wide variety of cellular lineages. Adipose tissue is from mesodermal origins and therefore, multipotent adipose-derived stromal cells will likely be most effective with application towards a mesodermal lineage. The ability to generate skeletal tissue is especially critical given the shortage of donor sites for an autograft and the inherent limitations of synthetic material including infection, r...

<table border="1">
 <tbody>
 <tr>
 <td>Name of the reagent:</td>
 <td>Company</td>
 <td>Catalogue number</td>
 <td>Comments (optional)</td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Lipoaspirate Harvest</td>
 </tr>
 <tr>
 <td>PBS</td>
 <td>Gibco</td>
 <td>10010-023</td>
 <td></td>
 </tr>
 <tr>
 <td>Hank's Balanced Salt Solution</td>
 <td>Cellgro</td>
 <td>21-023-CV</td>
 <td></td>
 </tr>
 <tr>
 <td>Collagenase</td>
 <td>Sigma</td>
 <td>C6885-500MG</td>
 <td></td>
 </tr>
 <tr>
 <td>Cell Strainer 100 &mu;m </td>
 <td>BD Falcon</td>
 <td>352360</td>
 <td></td>
 </tr>
 <tr>
 <td>Steri-top 500 ml .22 &mu;m filter </td>
 <td>Millipore</td>
 <td>SCGPT05RE</td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Calvarial Defect</td>
 </tr>
 <tr>
 <td>Z500 Brushless MicromotorsUM50C </td>
 <td>NSK </td>
 <td>NSKZ500</td>
 <td></td>
 </tr>
 <tr>
 <td>Circular Knife 4.0 mm </td>
 <td>Xemax Surgical </td>
 <td>CK40</td>
 <td></td>
 </tr>
 </tbody>
</table>

repair of a critical-sized calvarial defect model using adipose-derived stromal cells harvested from lipoaspirate

Craniofacial skeletal repair and regeneration offers the promise of de novo tissue formation through a cell-based approach utilizing stem cells. Adipose-derived stromal cells (ASCs) have proven to be an abundant source of multipotent stem cells capable of undergoing osteogenic, chondrogenic, adipogenic, and myogenic differentiation. Many studies have explored the osteogenic potential of these cells in vivo with the use of various scaffolding biomaterials for cellular delivery. It has been demonstrated that by utilizing an osteoconductive, hydroxyapatite-coated poly(lactic-co-glycolic acid) (HA-PLGA) scaffold seeded with ASCs, a critical-sized calvarial defect, a defect that is defined by its inability to undergo spontaneous healing over the lifetime of the animal, can be effectively show robust osseous regeneration. This in vivo model demonstrates the basis of translational approaches aimed to regenerate the bone tissue - the cellular component and biological matrix. This method serves as a model for the ultimate clinical application of a progenitor cell towards the repair of a specific tissue defect.

Watch this Scientific Journal Video about Repair of a Critical-sized Calvarial Defect Model Using Adipose-derived Stromal Cells Harvested from Lipoaspirate at JoVE.com

Repair of a Critical-sized Calvarial Defect Model Using Adipose-derived Stromal Cells Harvested from Lipoaspirate

This protocol describes the isolation of adipose-derived stromal cells from lipoaspirate and the creation of a 4 mm critical-sized calvarial defect to evaluate skeletal regeneration.

Craniofacial skeletal repair and regeneration offers the promise of de novo tissue formation through a cell-based approach utilizing stem cells. ...

repair-critical-sized-calvarial-defect-model-using-adipose-derived

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15.8K 조회수.  Stanford University. 이 프로토콜은의 고립을 설명 지방 - 파생 lipoaspirate과 골격 재생을 평가하는 4mm 중요한 크기 calvarial 결함의 생성의 stromal 세포를.