Stem Cell Technology Certificate Program

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Stem Cell Technology Certificate Program City
College of San Francisco Director Dr. Carin
Zimmerman Instructors Dr. Golnar Afshar Dr.
Shane Oram Dr. Jennifer Ng Dr. Alex
Zambon Dr. Carin Zimmerman
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• City College of San Francisco announces
• ground-breaking Stem Cell Research Technician
  Certificate Program
• SAN FRANCISCO, California, April 13, 2005
• San Francisco Mayor Gavin Newsom, City College
  of San Francisco Board President Rodel E.
  Rodis, Board Vice President Dr. Natalie Berg,
  and Chancellor Dr. Philip R. Day, Jr., announced
  today the receipt of a 779,067 grant under the
  California Governors Funding category, Growth
  Industries--High Wage, High Skill Job Training.
  This two-year grant will enable City College to
  create a
• Stem Cell Research Technician Certificate
  Program which will equip
• participants with the advanced skills needed to
  find jobs in the lucrative and rapidly growing
  field of stem cell research.

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WIA Workforce Investment Act
Benefits of WIA Job Seekers Universal access to
job search and labor market information Advice,
counseling, and support Education and skills
training Individual choice of service
Employers Influence over local area employment
policy Improved and trained employee pool
Development of on-the-job and customized
training opportunities Assistance for laid-off
workers Community Access to local area job
market information Improved workforce quality
Services designed for local area needs Reduced
need for welfare
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Stem Cell Technology Certificate Program

• How to get this program started?
• What skills are needed to prepare students
  (Undergraduate level) who
• complete the program for a career in Stem Cell
  Biology?
• What classes are needed to teach these skills?
• Establish collaborations with academic research
  institutions
• Dr. Mort Cowan - Director of Pediatric Bone
  Marrow Transplant (UCSF)
• Dr. Bruce Conklin - Gladstone Institute of
  Cardiovascular Disease (UCSF)
• Dr. Susan Fisher- Human Embryonic Stem Cell
  (UCSF)
• 4. Establish collaborations with industry--
  Miltenyi Biotec
• 5. Set up internship program for students who
  complete the programFisher Lab UCSF

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Stem Cell Technology Certificate Program
3 Courses BTEC21A Mammalian Cell Culture (6
weeks/9 hours per week) BTEC21B Fluorescent Cell
Technology (6 weeks/9 hours per week) BTEC21C
Stem Cell Technology (8 weeks/9 hours per
week) BTEC24 25 Introductory and Advanced PCR
BTEC22 ELISA BTEC23 Western Blotting Minimum
prerequisites for BTEC 21A One semester of
college-level general biology with lab and two
semesters of college-level general Chemistry
including organic with lab. Required
Texts Culture of Animal Cells A manual of basic
technique (R. Ian Freshney, 5thEdition)
Courses offered in the Fall, Spring, and
Summer. Day and Night Classes. Students with
proper background are capable of completing
program in 1 calendar year.
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CCSF Cell Culture Laboratory/Classroom

• Our basement space functions as a classroom.
• And as a laboratory.

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Cell Culture Laboratory

• We have 4 CO2 incubators for growing and
  maintaining cells.
• We have 6 laminar flow hoods--some of them are
  double hoods providing 10 workspaces for the
  students.

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Cell Culture Laboratory

• We have 2 FACScan for stem cell
  isolation/analysis.
• We have 6 epifluorescence inverted microscopes,
  as well as regular light inverted microscopes.

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BTEC21AMammalian Cell Culture
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Cell culture techniques

• Cell culture equipment
• Operation and maintenance
• Plating and feeding cells
• Defrost and freeze and proper storage of cells
• Counting cells
• Hemacytometer and coulter counter methods
• Trypsinizing and subculturing
• Media components and preparation
• Staining and counting colonies
• Calculate cell concentrations
• For appropriate cell seeding, transfer and
  feeding

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Cell culture concepts

• Understanding and managing contamination
• Cell cycle and cell death
• Plating efficiency
• Cell concentration and density
• Doubling time
• Phases of growth cycle
• DNA damage and repair
• Genetic disorders (Xeroderma pigmentosum)
• Viability, toxicity and survival

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Laboratory exercises

• Plating efficiency
• CHO and COS-7 cells
• Growth rate (colony size) cell survival (colony
  number)
• Data collection and analysis

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• Phases of Growth Cycle
• Lag, log and plateau phases
• Saturation density
• Population doubling time
• Medium volume, cell concentration and cell
  density
• Data collection and analysis

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Cytotoxicity Assay

• Model Xeroderma Pigmentosum
• Cells AA8 (wild type), UV5 (NER deficient) and
  5P2E
• (restored NER by transfection)
• Toxic agent UV light
• Viability assay

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Survival (clonogenic) assay
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Skills

• Reading (handouts papers)
• Writing (Laboratory notebooks)
• Designing and performing experiments
• Observing and collecting data
• Calculation and analysis
• Reasoning and problem solving
• Working with others and basic laboratory
  etiquette

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BTEC21BFluorescent Cell Technology
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BTEC21B Fluorescent Cell Technology (6 weeks/ 9
hours per week)
MODEL Additional studies with DNA repair
pathways Cell cycle control and apoptosis, cancer
biology Reporter gene assays Gene Therapy Gene
expression and transfection of mammalian
cells CHO (AA8, UV5) and COS7 cells SKILLS Descr
ibe different types and applications of
fluorophores used in cell biology research.
Demonstrate proper use and care of a
fluorescent microscope. Perform nucleic acid
staining with PI , Hoescht Perform live/dead
cell staining assay with PI and Calcein AM
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BTEC21B Fluorescent Cell Technology(6 weeks/9
hours per week)
SKILLS (Continued) Perform organelle-specific
staining with various fluorophores Perform
transfections using fluorescent markers Perform
gene therapy transfections to rescue a mutant
cell line. Analyze transfected cells using
fluorescent microscopy. Identify subcellular
localization MitoTracker, LysoTracker,
GFP-actin, etc. Perform immunostaining and
apoptosis assays. Discuss current applications
of fluorescent technology, DNA microarrays,
confocal microscopy and FACS analysis.
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BTEC 21C Stem Cell Technology
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BTEC21C Stem Cell Technology (8 weeks/ 9 hours
per week)
LECTURE COMPONENT Ethics of stem cell biology
and cloning Introduction to stem cell biology,
self renewal, and pluripotency ES cells and
primordial germ cells in early development Adult
stem cells and research applications
Hematopoietic stem cells, mesenchymal stem
cells and bone marrow transplants Engineering
stem cells and knockout mice Uses of stem cells
in regenerative medicine amniotic fluid stem
cells chord blood stem cells RNAi techniques in
stem cells
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BTEC21C Stem Cell Technology (8 weeks/ 9 hours
per week)
LABORATORY COMPONENT 1) Isolation of stem cells
from adult mouse Dr. Mort Cowan Director of
Pediatric Bone Marrow Transplant
(UCSF) Isolation of hematopoietic and
mesenchymal cells from mouse bone marrow
(Miltenyi Biotec Products) 2) Introduction and
basics of flow cytometry Analysis of
cells using a FACScan 3) Culturing mouse ES
cells Dr. Bruce Conklin Gladstone Institute
of Cardiovascular Disease (UCSF) Growing feeder
cells and ES cells while maintaining their
pluripotency 4) Preparing ES cell karyotypes
Preparing/Analyzing chromosome spreads 5)
Differentiation of ES stem cells Hanging
drop technique for differentiation of ES
cells Use of antibodies and immunostaining to
confirm differentiation of ES cells
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BTEC21C Stem Cell Technology (8 weeks/ 9 hours
per week)
SKILLS Relate principles of stem cell biology
to proper laboratory techniques Critically and
independently assess the reality and future of
applied stem cell biology Differentiate between
different types of stem cells in terms of their
function, use and care Isolate mesenchymal and
hematopoietic stem cells from mouse bone
marrow Isolate c-kit positive hematopoietic stem
cells Describe the underlying principles of flow
cytometry Use a FACScan (flow cytometer) Analyse
cell surface markers and cell types using a
FACScan
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BTEC21C Stem Cell Technology (8 weeks/ 9 hours
per week)
SKILLS (Continued) Handle and care for
embryonic stem (ES) cells while maintaining
their pluripotency. Properly culture and
maintain feeder cells. Use proper culture
techniques to induce differentiation of feeder
Independent ES cells into different cell
types. Perform karyotyping on ES
cells. Properly freeze and thaw stem cells to
maintain viability. RNAi and engineering of stem
cells.
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Internships

• Collaborate with Dr. Susan Fisher at UCSF.
• Students who successfully complete their
  certificate participate in a 3-4day training with
  human ES cells at UCSF.
• 12 students went through the training this past
  December/January, with another group to be
  scheduled in the fall semester.

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Mouse ES Cells Differentiated Into Beating
Cardiac Myocytes
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Future Stem Cell Technicians at Work!
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Stem Cell Certificate Success!

• To date, we have had 10 BTEC21C classes, with
  over 100 students receiving their certificates.
• Many of our graduates have been hired for cell
  culture work at UCSF, Stanford, Gladstone
  Institutes, Stem Cell Inc., Progenitor,
  Genentech, etc.
• With CIRM grant funding on the horizon, we
  estimate many more positions will become
  available to our graduates.

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Acknowledgements
CCSF Phil Jardim Kristin Hershbell Dr. Carin
Zimmerman Dr. Golnar Afshar Dr. Alex
Zambon Dr. Jennifer Ng Dr. Edith Kaeuper Dr.
Wing Tsao Dr. Carole Toebe Dr. Elaine Johnson
UCSF/Gladstone Dr. Mort Cowan Ms. Elizabeth
Dunn Dr. Bruce Conklin Dr. Jennifer Ng Dr.
Susan Fisher
Miltenyi Biotech Mr. Thad Baker Ms. Julie
Rae Dr. Shane Oram