Starting a Stem Cell Lab

A stem cell lab is usually like any standard cell culture research lab. Significant considerations to establish a stem cell lab are:

I. Good lab design

The design of the stem cell lab is similar to any other animal cell culture lab. The team usually consisted of an architect, contractor, builder, plumber, and laboratory director, and costs usually cost 3-4 times that of office space Key considerations of the lab design are:

Space:
  • Includes the number of personnel working at a time
  • Major equipment footprint
  • Storage room facility
  • Other major functional areas (e.g., tissue and cell culture)

Scope of work – biosafety level of lab

Approval of regulatory bodies- In India, this may not be applicable. However, in other countries like the USA, culturing human stem cells may require permissions, including academic institutions, like the Stem Cell Research Oversight Committee, Institutional Review Board (IRB), Biosafety Committee, and Institutional Animal Care and Use Committee (IACUC).

II. Quality control –

Quality controls (QCs) include some basic procedures and protocols related to the methodology of work with highest standards keeping safety as priority. Like Standard operating procedures (SOPs), validated reagents, reliable techniques, and checklists. These are discussed below in brief.

III. A standard operating procedure (SOP)

is a set of step-by-step instructions which are made to help students or employees to work in a lab/place safely. Each lab has different SOPs depending on the type of research carried out in particular laboratories.

Validated reagents are as follows which may vary to different labs depending on their scope of work

  • Media (low glucose containing DMEM or DMEM-F12)
  • Media (high glucose containing DMEM or DMEM-F12)
  • MSC and/or HSC commercial media (e.g., StemSpan)
  • MSC and/or HSC commercial growth media growth factor supplements
  • Fetal bovine serum (FBS)
  • Sodium pyruvate solution
  • Penicillin-streptomycin (Lonza or Gibco)
  • Dulbecco's PBS without calcium and magnesium (D-PBS)
  • Hank's blank salt solution (HBSS)
  • Trypsin for cell detachment
  • Collagenase type I or type IV – for enzymatic digestion of tissue from which stem cells are to be harvested.
  • Trypan Blue solution (0.4%) – live and dead cell counting
  • 0.5% Triton X-100
  • Bovine serum albumin (BSA) – blocking buffer
  • Normal goat serum–blocking buffer for primary antibodies
  • Primary antibodies for stem cell markers– Species-specific. All mouse-specific antibodies may not work in canines or bovines or vice versa
  • Trizol or RNAiso reagent (Ambion or Takara) – for RNA isolation for validation of stem cell-specific transcripts
  • Ultrapure DEPC treated or nuclease-free water
  • Ethanol – for cleaning of laminar flow
  • 3% hydrogen peroxide
  • RIPA buffer – for protein isolation and validation of stem cell protein biomarkers
  • Cell freezing media

IV. Skilled technical personnel:

A person familiar with lab protocols and having rich experience in managing a lab is required to keep track of the things going on in the lab. These people may require to perform all necessary jobs associated with the lab, including providing basic lab safety training to new students and visitors.

Funding resource: Financial support for running a lab mostly comes from the grant money, which can be utilized to order consumables, reagents, manpower, etc.

VI. Necessary equipment/machines:

Some important standard equipment required for a stem cell lab are mentioned below

  • Biosafety cabinet class II
  • CO2 incubator
  • Aspirator device
  • Water bath
  • Hemocytometer – for manual counting and calculation of live and dead cells. However, automated cell counters like Countess 3 Automated Cell Counters (Invitrogen) and TC20 Automated Cell Counter (BioRad) is recommended for rapid and precise cell counting.
  • Refrigerated Centrifuge – low speed for spinning of cells
  • Multi-speed Eppendorf centrifuge
  • Vortex and tube shakers
  • Pipettors
  • 15 mL, 50 mL conical tubes (Corning or Axygen)
  • Syringes and needle (B.D. science)
  • 70 µM and 40 µm pore size nylon mesh filter (Corning or B.D. Science)
  • Microtubes (1.5 mL; ThermoFisher or Tarson)
  • Cryovials (1.5 mL Corning)
  • Tissue culture dishes (100 mm)
  • Tissue culture flasks (75 cm2 and 25 cm2)
  • Tissue culture plates (96-well, 48-well, 24-well and 6-well)
  • CoolCell LX freezing container or Mr. Frosty
  • Phase contrast microscope
  • Storage
    • Cabinets and shelves
    • Refrigerators (2-8 °C)
    • Freezer (-20 °C)
    • Deep freezer (-80 ºC)
    • Cryofreezer – (-130 ºC or liquid nitrogen)
  • Optional equipment
    • RT-qPCR
    • Flow cytometer
    • Automated electronic cell counter - recommended

VII. Define the scope of work:

Stem cell research is focused on studying and understanding stem cells, their nature, and their mechanism of action by analyzing various pathways involved. Research on stem cells reveals their vast potential in treating many incurable diseases in human and veterinary medicine.

VIII. Major specific functional areas:

According to the expertise of the project investigator (PI)

  • Storage room
  • Cold room
  • Cryo-storage
  • Biosafety for human materials handling
  • Institutional approvals for animal and or human subjects’ investigations (e.g., ethical practice)
  • Types of stem cells and number of cell types to be used at a time

IX. The number of personnel working-

determines the space area, availability of equipment, and storage of incoming samples and stem cell products produced.