1. HYALURONIC ACID-BASED SCAFFOLD FOR GROWING HUMAN ADIPOSE-DERIVED STEM CELLS IN IMMUNODEFICIENT MICE
FENG-SHU CHANG, SIN-DAW LIN, SHU-HUNG HUANG,
TSAI-MING LIN, YUN-NAN LIN, CHIA-CHENG CHANG,
CHEE-YIN CHAI, HSUEH-WEI CHANG
DEPARTMENT OF PLASTIC SURGERY
KAOHSIUNG MEDICAL UNIVERSITY CHUNG-HO MEMORIAL HOSPITAL
KAOHSIUNG TAIWAN, REPUBLIC OF CHINA
“NOTHING TO DISCLOSE.”

2. To discover an ideal filler:
The goal of the study
To discover an ideal filler:
Long lasting
Safe
Effective
Biocompatible
Low immunogenicity
 Human Adipose-Derived Stem Cells ( hASCs) with hyaluronic acid get.

3. MATERIAL AND METHODS
For in vitro studies:
hASCs: isolated from human adipose tissue and were confirmed by multi- lineage differentiation and flow cytometry.
hASCs were cultured on HA gel
Effectiveness of cell attachment and proliferation on HA gel was surveyed by inverted light microscope.

4. 8 weeks post injection implants were harvested for analysis
MATERIAL AND METHODS
For in vivo studies: fillers were injected subcutaneously into back to nude mice (n=6) in each group
HA gel containing hASCs
hASCs without HA gel
HA gel alone
8 weeks post injection implants were harvested for analysis

5. RESULTS
The hASCs were well-differentiated into three cell lineage.

6. RESULTS
Figure 2. Inverted light microscope image showing hASCs seeding in HA gel. hASCs proliferate and aggregate in HA gel on Day 1, 3, 7 (X100) (red color indicated hASCs with CM-Dil stain). An MTT assay was used to check hASCs cell proliferation in HA gel. No significant difference between HA-hASCS and hASCs alone group in MTT assay.

7. RESULT
Figure 3. Macroscopic appearance of grafts in nude mice. Suspension of HA gel containing hASC was subcutaneously injected into the back of each nude mouse. (Left) New adipose tissue formation was noted near the residual HA gel at eight weeks after injection. (Right) In the control group, the residual clear HA gel was noted in subcutaneous areas. Ruler = 2cm.

8. RESULTS
Figure 4. Histological examination of the grafts stained by HE stain eight weeks after injection of suspension of HA gel with hASCs revealed newly formed adipose tissue. Black stars (*) indicate adipose tissue (scale bar 50 μm). Oil-Red stain (red arrows) revealed the gradual replacement of degraded HA gel with newly formed adipose tissue (scale bar 100 μm). In the immunohistochemical stain showed CD44 positive cell marker and CM-Dil positive in newly adipose tissue (scale bar 50 μm, 100 μm).

9. RESULT
Figure 5. Results of PCR analysis of Alu gene in graft specimens.
1) HA gel + hASCs grafts.
2) HA gel only.
3) hASCs only.
4) HA gel with medium.
5) control group.
P was positive control.

10. A permanent effect of an injected filler is possible:
CONCLUSION
A permanent effect of an injected filler is possible:
the volume of HA gel is replaced by new adipose tissue generated by hASCs
A promising approach for developing long lasting filler.

11. Thanks for your attention！
K.M.U. TW.

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