Core Facility for Human Pluripotent Stem Cell Research

VISIONS & MISSIONS:

Techniques of manipulating human pluripotent stem cell (hPSC) in culture can be challenging.  The core is dedicated to

  1. establishing the state-of-the-art technology platforms in University of Hong Kong (HKU) / Li Dak-Sum (LDS) Research Centre to support stem cell research
  2. providing consultation and technical support to conduct research with hPSC lines
  3. helping researchers learn techniques that can be used later to initiate stem cell research in their own labs 
  4. With a long-term aim to fostering collaborations and interactions between researchers and laboratories within HKU and around Hong Kong. 

CORE SERVICES:

Core Services:

Human Pluripotent Stem Cell Culture Service:
    • establishing induced pluripotent stem cell lines
    • characterizing established human pluripotent stem cell lines
    • monitoring previously established hPSC lines
        o Karyotyping
        o Mycoplasma testing
    • providing cryostore service for established hPSC lines
    • providing training for those wishing to utilize hPSC as a model system

Stem Cell Genomic Service:
    • generating genetic modified hPSC lines
    • providing training for utilizing CRISPR/Cas9 system on hPSC 
    • providing consultation service for the construction of donor plasmid and selection of gene-targeting platforms

Stem Cell Reagent Core:
    • plasmids for reprogramming
    • plasmids for gene-editing 
    • control hPSC lines
    • culture reagents for hPSC culture and differentiation

Others:
    • single cell sorting compatible to single cell culturing or single cell RNA sequencing
    • consultation service for grant application
    • advanced or customized services are available in collaboration basis


CORE TEAM MEMEBERS

The team is led by Dr. Elly Ngan, Associate Professor in Department of Surgery.  Dr. Ngan is a core principal investigator in several collaborative research grants for stem cell research: SK Yee grant (HK$5M), a RGC funded Theme-based Research Scheme (TRS) project (HK$62.36M) and a ITF funded UICP project (HK$7.6M).  More recently, Dr. Ngan was awarded a Seed Funding (HK$3M) under the impactful scheme from LDS Research Centre for the establishment of high-throughput CRISPR screening and human intestinal organoid platforms.  


With the support of these collaborative grants, the team has acquired the state-of-the-art technologies for stem cell research from the world’s leading research institutes including Center for iPS Cell Research and Application (CiRA), Harvard Stem Cell Institute and the Cincinnati University, etc.  In the past few years, the team has established various technology platforms and facilities to support the stem cell research in HKU. 


Two senior scientists are overseeing the core facilities and services:

  1.  Dr. Frank Lai is a senior scientist and has substantial experience in molecular biology, gene-targeting with mouse ES and human PSC. He is in charge of the development of CRISPR/Cas9 platforms.  He will provide technical support on utilization of genome-editing platform on hPSC and generation of genome-edited hPSC.
  2. Dr. Cynthia Lau is a senior scientist and has substantial experience in human PSC culture and differentiation. She is in charge of the development of reprogramming platform and organoid culture.  She will provide technical support on hPSC culture, maintenance and differentiation.

TEAM’S EXPERTISE:

Episomal reprogramming (follows the protocol established by Shinya Yamanaka, CiRA, Kyoto University)
Maintenance and characterization of hPSC
Monitoring the genomic integrity of hPSC
Utilization of CRISPR/Cas9 system on hPSC (Protocols provided by Chad Cowan, the Harvard Stem Cell Institute) for generation of
        • Deletion/truncation
        • Base-editing
        • Mutation (through oligo or donor plasmid)
        • CRISPR-mediated gene activation
        • Multiplex targeting
Single cell sorting and culturing of hPSC
Single cell sequencing
ATACseq
Derivation of CNS and PNS neurons, mesenchymal stem cells, smooth muscle cells, chondrocytes and osteocytes from hPSC
Generation of intestinal organoids with nervous system using hPSC (Protocols provided by Prof James Wells group, the University of Cincinnati) 
Physiological characterization of muscle strip
Analysis of cell migration with live imaging 
Confocal imaging 


CORE EQUIPMENTS:

BDAriaII
BDAriaIII
Microscopes:
Nikon DS-Qi2 (full-frame COMS) and DS-U3 camera for fluorescent and bright-field image
Leica MZ10 F Stereomicroscope with fluorescence (3 colors)
Leica S8APO Stereomicroscope
EVOS XL Core
Bioreactor for 3D organoid culture
PerkinElmer Victor X4 Multi-label Microplate Reader with fluorescence filters
Cell Counter “Countess II FL”
Leica CM3050 S Research Cryostat
DMT 820MS Muscle Strip System
Warner PLI-90A Plus Pico Injector Plus
CO2 incubators
TC hoods


SERVICE CHARGES:

Services

Unit Price (HKD)

Contact person

Email address

Mycoplasma test

$1,000

Dr. Cynthia Lau

cynlau@hku.hk

gRNA construct generation

$8,000-$12,000

Dr. Frank Lai

laifrank@hku.hk

Cas9 Genome editing

$8,000-$15,000

Dr. Cynthia Lau

cynlau@hku.hk

Custom services

To be determined

Dr. Elly Ngan

engan@hku.hk

General enquiries

-

Dr. Thomas Leon

tleon@hku.hk










Remarks:

Specific terms and conditions apply to the above services and please contact us for the details. All prices are subject to change.


CONTACT US:


 

Name

Contact information

Team Leader

Dr. Elly Ngan

engan@hku.hk

Senior Scientist

Stem cell genomics core

Dr. Frank Lai

laifrank@hku.hk

Senior Scientist

Stem cell culture core

Dr. Cynthia Lau

cynlau@hku.hk

Manager

Dr. Thomas Leon

tleon@hku.hk