분회초청강연

     나노바이오

  김태형 교수 중앙대학교

     ■ Education

■  Academic Posisition

Label-free and non-destructive characterizations of stem cell differentiation 

and organoid generation

Stem cells have emerged as valuable sources for regenerative therapy and drug screening. They can generate multiple types of cells and are capable of forming an organ-like structure in vitro that is named organoids. However, precise characterizations of the stem cell differentiation into specific lineages and organoids are still challenging without loss of cell viability and function. In this study, we report a new type of analytical platform that enables precise, non-destructive, and label-free characterizations of several key stem cell feature including pluripotency/multipotency, differentiation and organoid maturation.

The autofluorescence and Raman integration (ARMI) method was developed as an optical tool for the rapid analysis of adipogenic and osteogenic differentiation of human adipose-derived mesenchymal stem cells (hAMSCs) in vitro. The method was further advanced to a three-dimensional analysis of odontogenesis of human dental pulp stem cells (hDPSCs) spheroids based on the β-carotene and HA depositions.

Besides the optical platforms, the electrochemical approach was also developed as an efficient tool to selectively detect the undifferentiated human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). The remaining stem cells were successfully quantified using a Geltrex-modified highly conductive gold nanostructure (HCGN) platform after kidney organoid generation. The Same approach was also shown to be applicable to cancer cell research where the HCGN effective in generating multicellular cancer cell spheroids and viability detection under drug treatment conditions.

Therefore, it can be concluded that our platform is highly useful as a new label-free and non-destructive tool for stem cell characterizations, which will ultimately contribute to the development of regenerative medicine.

  박성수 교수 성균관대학교

     ■ Education

■  Academic Posisition

Tumor microenvironment recapitulated by microfluidics and 3D bioprinting

Thanks to advances in microfabricated cell culture and 3D bioprinting technology over the past decades, the ability to realistically simulate important biological steps in disease models including cancer has dramatically improved. In this presentation, I will show our recent efforts to recapitulate tumor niches displaying angiogenesis and invasion using 3D bioprinting and microfluidic devices. For an example, cancer drug resistance accelerator chip, consisting of approximately 500 hexagonal micro-compartments with antiparallel gradients of doxorubicin and cell nutrients, can rapidly induce drug resistance in U87 glioblastoma cells. Another example is to print 3D tumor composed of fibroblasts, endothelial cells and breast cancer cells recapitulating angiogenesis. These works demonstrate that miniaturized cell culture system is an alternative to existing 2D culture models for cell biologists as well as medical researchers who are interested in understanding complex cell behaviors in an in-vivo like condition.

  송대섭 교수 고려대학교

     ■ Education

■  Professional Record

Nanobiotechnology for diagnosis, vaccine and treatment against Disease X

; unexpected emerging viral diseases

Unlike in the past, emergences of viral infections have been and are occurring at very frequent intervals, causing enormous deaths and disability worldwide. Against a constant background of established infections, periodical emerging the epidemics of highly pathogenic influenza viruses (HPAI) greatly expand the global burden of infections. Accurate and rapid diagnosis of viral infections can result in effective and appropriate prevention and quarantine measures. This study compares and discusses about rapid diagnostics for the differential patho-typing between HPAI and LPAI using nanobiotechnology. The field of nanotechnology encompasses those technologies to fabricate materials, including sphere, cubic and nanoscale particles. Therefore, nanobiotechnology has the potential to offer not merely advances in diagnostics and vaccination to control infectious diseases but also in delivering various capabilities as below; Highly pathogenic avian influenza virus (HPAIV) infections have occurred continuously and crossed the species barrier to humans, leading to fatalities. A PCR-based molecular test is currently the most sensitive diagnostic tool for HPAIV; however, the results must be analyzed in centralized diagnosis systems by a trained individual. This requirement leads to delays in quarantine and isolation. To control the spread of HPAIV, rapid and accurate diagnostics suitable for field testing are needed, and the tests must facilitate a differential diagnosis between HPAIV and low pathogenic avian influenza virus (LPAIV), which undergo cleavage specifically by trypsin- or furin-like proteases, respectively. In this study, we have developed a differential avian influenza virus (AIV) rapid test kit and evaluated it in vitro and using clinical specimens from HPAIV H5N1-infected dogs. We demonstrated that this rapid test kit provides highly sensitive and specific detection of HPAIV and LPAIV and is thus a useful field diagnostic tool for H5N1 HPAIV outbreaks and for rapid quarantine control of the disease. In addition to diagnostics, development of better adjuvant accompanied with vaccine for enhancing immunogenicity has been greatly required for the control of influenza infection. Herein, we also address nano-complex of amphiphilic grafted poly (amino acid) and hydrophobic squalene (PA/S-NC) as a potent adjuvant that can act as a robust strategy for induce humoral (Th2) and cellular (Th1) immune responses as well as a delivery agent of antigens. CASq performed great biocompatibility, particle stability, and produced a high degree of antigen-specific antibodies and T cell immune responses in mice when CASq was co-administered with inactivated whole influenza virus antigen (CA04), in which CASq exhibited complete protection against lethal infection.

Keyword: HPAIV, nanobiotechnology, diagnostics, adjuvant, vaccination

  김중배 교수 고려대학교

     ■ Education

■  Professional Record

Nanobiocatalysis and its potential applications

Enzymes are biomolecules that can catalyze a variety of chemical reactions. However, their practical applications are often hampered by their poor stability. Nanobiocatalysis, immobilizing enzymes using various nanomaterials, has demonstrated its successes in stabilizing the enzyme activity for various enzyme applications such as biosensors, biofuel cells, enzyme-linked immunosorbent assay, membrane antifouling, and CO₂ conversion. This presentation will cover nanobiocatalytic stabilization of acylase and carbonic anhydrase for their successful uses in enzyme-catalyzed quorum quenching and CO₂ conversion, respectively. Especially, the stabilization of carbonic anhydrase has made an unprecedented success by maintaining 83% of initial enzyme activity even after incubation in aqueous solution under shaking at 200 rpm for two years. Stabilized carbonic anhydrase was successfully employed for the effective conversion of CO₂ to bicarbonate, which was further used for expedited microalgae growth and improved calcium carbonate production. If time permits, several other examples of nanobiocatalytic stabilization and applications will also be introduced.

  이성운 대표 (주)레보스케치

     ■ Education

■  Work History

Next Generation Real-time Digital PCR

 중합효소연쇄반응(PCR; Polymerase Chain Reaction)은 1984년 발명된 이후에 발전을 거듭하여 현재 3세대 Digital PCR 기술로 빠르게 발전해 왔다. 기존 기술에 비해 비약적인 민감도 성능 개선이 이루어졌지만 3~4개 장비로 이루어지고 증폭 후에 한번만 형광을 측정하는 End-point Scan을 사용함으로써 사용이 불편하며 결과가 부정확해지는 문제를 가지고 있는 상태이다. 이에 RevoSketch는 하나의 Digital PCR 기기에 필요한 모든 기능을 통합하였으며 최초로 증폭 주기마다 스캔이 가능한 Real-time Digital PCR 기술을 개발하였다. 또한, 원심력을 이용한 자동 분주 구조와 Nano-imprinting 기법을 이용한 경제적인 소모품 생산 기술을 개발하였다. 결론적으로, 비전문가의 편리한 사용이 가능한 고 민감도 성능을 제공하는 동시에 대량 생산이 가능한 저비용 소모품 구조로 경제적인 Digital PCR기술을 제공 한다.