줄기세포와 오가노이드 3D 배양을 위한 Biosilk | 고마바이오텍

특징

• Biosilk는 spidroin (스피드로인)이라고 하는 거미줄 단백질 성분을 이용한 3D culture matrix 입니다.
• 세포외기질(ECM)과 유사한 네트워크를 형성하여 실제 조직 환경을 모사합니다. 
  
• Silk fiber가 형성되는 동안 생기는 air bubble의 결과로 porous하여 media의 영양분과 산소를 훨씬 더 많이 접촉할 수 있습니다.  
• Biocompatible, non-immunogenic, biodegradable합니다.
• 성분이 규명되어 있으며, 동물성분이 포함되어 있지 않습니다. 
  
• 오가노이드 배양시, necrotic center가 생성되지 않습니다.
• Biosilk와 Biolaminin 521이 혼합된 Biosilk 521은 hESC 및 iPSC의 증식과 분화를 위한 3D 배양 환경을 조성합니다. 

Representative pictures of the Biosilk 521 foam taken from above and from the side at A) day 0, B) day 1 and C) day 3 after cell seeding. The bubbles have dispersed after 3-5 days, transforming the foam into a Biosilk network (D) with evenly integrated cells (nuclei staining; DAPI) attached to the microfibers (E). 

Biosilk를 이용한 세포/세포주

Primary cells

• HSkMSC – human skeletal muscle satellite cells
• HDMEC – human dermal microvascular endothelial
• Schwann – human Schwann cells
• HMSC – human mesenchymal stem cells
• HDF – human dermal fibroblasts
• MDMEC – mouse dermal microvascular endothelial
• MMSC – mouse mesenchymal stem cells
• HUVECs – human umbilical vein endothelial cells

Cell lines

• HaCaT – Human keratinocyte cell line
• MIN6m9 – insulin-secreting mouse pancreatic β-cell line
• MCF7 – human breast cancer cell line

Pluripotent stem cell lines

• HS980 – human embryonic stem cell line
• HS975 – human embryonic stem cell line
• C5 – human induced pluripotent cell line
• iPSC3 – human induced pluripotent cell line

데이터

A) Bright-field images of ventral midbrain organoids with Biosilk (silk-VM org) display less distinct outer and innner parts compared to conventional organoids (VM org) at day 12. Scale bar 200 μm. B) Biosilk organoids show no necrotic centers after 6 months of culture in contrast to conventional organoids. TUNEL staining, scale bar 100 μm.

The Biosilk foam looks almost transparent at day 1(IV) and denser after two weeks (V). The 3D network is filled with well-spread stem cells (blue) already after 3 days (VI). Modified from Figure 1in Johansson et al. 2019 Scientific Reports licensed under CC BY 4.0.

Micrographs of hPSCs attached to Biosilk 521 days 2, 3, and 5 after seeding. The cells are evenly integrated between the Biosilk microfibers. 

Differentiation of multipotent adult stem cells. Human mesenchymal stem cells (HMSC) in Biosilk foam differentiated into the adipogenic lineage containing lipids, visualized by Red Oil staining (red). Scale bar = 100 µm. Adapted from Figure5in Johansson et al. 2019 Scientific Reports licensed under CC BY 4.0.