Supporting Information

Injectable and Detachable Heparin-Based Hydrogel Micropatches for Hepatic Differentiation of hADSCs and Their Liver Targeted Delivery

YoungMin Hwanga1, MeeiChyn Goha2, Mihye Kima3, Giyoong Taea*

a School of Materials Science and Engineering, Gwangju Institute of Science and Technology, 123 Cheomdan-gwagiro, Buk-gu, Gwangju 61005, Republic of Korea.

*Corresponding authors: Giyoong Tae

School of Materials Science and Engineering, Gwangju Institute of Science and Technology, 123 Cheomdan-gwagiro, Buk-gu, Gwangju 61005, Republic of Korea.

Tel: +82-62-715-2305

Fax: +82-62-715-2304

Email: gytae@gist.ac.kr

Formation of polyelectrolyte multilayer (PEM),(PLL/HA)2-(GMA-chi) on indium tin oxide

(ITO) substrate:

The ITO electrodes was cleaned by ultrasonication in isopropanol and deionized water for 10

min each prior to the polyelectrolyte absorption. Then, the electrodes were dried with filtered

nitrogen, followed by an oxygen plasma (YES-R3, San Jose, CA, USA) treatment at 300 W for 5

min.A total 2 layers of poly (ʟ-lysine) (PLL) and hyaluronic acid (HA) (PLL/HA) coated on the

substrate were done by immersing the substrate in PLL (0.5 mg/ mL) and HA (0.5 mg/ mL)

polyelectrolyte solution for 5 min each and followed by rinsing with PBS buffer after each step.

A top layer of glycidyl methacrylated chitosan (GMA-chi) was coated by immersing the

substrate in a GMA-chi (30% acrylation, 10 mg/mL) solution for 30 min.

Human nuclei immunofluorescence staining

Livers of rats were dissected after 5 and 15 days upon intravenous injection of DiD-labeled

hepatic-like cells. The liver samples were fixed in 10 % formalin solution and paraffin-

embedded. 5 m-sections were deparaffinized and followed by antigen retrieval in 0.25 %

Triton-X100 solution (in PBS) for 15 min at 37 °C. Non-specific antibody binding was prevented

via incubation for 1 h at room temperature with 1 % BSA. Sections were washed and incubated

with PBS-diluted human nuclei antibody (1:100 mouse monoclonal antibody to human nuclei,

Millipore, Billerica, MA, USA) overnight at 4 °C. Sections were washed and incubated with

FITC goat polyclonal antibody to mouse (Santa Cruz, Dallas, Texas, USA, 1:100 in PBS) for 1 h

at room temperature. Sectioned were washed in PBS and mounted with 4’, 6-diamidino-2-

phenylindole (SlowFade® Gold Antifade Reagent with DAPI, Life Technologies, Grand Island,

NY, USA). Final sections were visualized by fluorescence microscopy (DFC 450, Leica,

Wetzlar, Germany).

Figure S1. Degradation rate of heparin-based hydrogel with various concentrations in PBS.

Figure S2. Human nuclei staining of paraffin liver section after 5 and 15 days upon intravenous

injection of DiD-labeled hepatic-like cells. (Human nuclei: green; Nuclei: blue). Scale bar: 200

µm

Figure S3.Quantification of fluorescence intensities of four organs (liver, lung, spleen and heart)

of rats after 1 day upon intravenous injection with DiD-labeled hepatic-like cells. Data are

expressed as means ± STD with n=3. *p<0.05.