Tulane hMSC Protocol

Question 1

hMSC Acronym

Answer 1

Human Mesenchymal Stem/Stromal Cell

Question 2

Reagent

Answer 2

a substance or mixture for use in chemical analysis or other reactions.

Question 3

Stock Solution

Answer 3

a concentrated solution that will be diluted to some lower concentration for actual use.

Question 4

CFU

Answer 4

Colony Forming Unit

Question 5

Assay

Answer 5

an investigative (analytic) procedure in laboratory medicine, pharmacology, environmental biology and molecular biology for qualitatively assessing or quantitatively measuring the presence, amount, or functional activity of a target entity (the analyte).

Question 6

Phase Contrast Microscopy

Answer 6

an optical microscopy technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image. Phase shifts themselves are invisible, but become visible when shown as brightness variations.

Question 7

Incubator Temperature and CO2 Levels

Answer 7

37° Celsius, 5% CO2

Question 8

Long Term Storage Temperature

Answer 8

-180° Celsius

Question 9

RS Cells

Answer 9

Rapidly Self-Renewing Cells

Question 10

Proliferate

Answer 10

increase rapidly in numbers; multiply.

Question 11

Senescence

Answer 11

the condition or process of deterioration with age; loss of a cell’s power of division and growth.

Question 12

All media should be warmed to what temperature before use?

Answer 12

37° Celsius

Question 13

Before you bring ANYTHING under the hood, what should you do?

Answer 13

Spray the object with 70% ethanol to reduce the risk of contamination

Question 14

What do you ALWAYS wear while working with the cells?

Answer 14

A lab coat and gloves to reduce the risk of contamination

Question 15

Reagent (Cell Culture): αMEM with L-glutamine

Answer 15

Alpha Minimum Essential medium with L-glutamine, but without ribonucleosides or deoxyribonucleosides

Question 16

Reagent (Cell Culture): FBS

Answer 16

Fetal Bovine Serum, premium select, hybridoma qualified, not heat inactivated

Question 17

Reagent (Cell Culture): L-glutamine

Answer 17

(200mM) in solution of 0.85% NaCl

Question 18

Reagent (Cell Culture): PS

Answer 18

Penicillin G (10,000 units/ml) and streptomycin sulfate (10,000 micrograms/ml) in solution of 0.85% NaCl

Question 19

Reagent (Cell Culture): PBS

Answer 19

Phosphate Buffered Saline, 1X. Used to wash the flask and cells. DO NOT USE WATER, or the cells will burst from changes in the osmotic pressure and pH.

Question 20

Reagent (Cell Culture): Trypsin/EDTA

Answer 20

0.25% Trypsin and 1mM ethylene diamine tetraacetic acid (EDTA) in Hanks’ Balanced Salt Solution. This reagent will lift the adhered cells.

Question 21

Reagent (Cell Culture): Trypan Blue

Answer 21

0.4% Trypan Blue in solution of 0.85% NaCl

Question 22

Reagent (Cell Culture): DMSO

Answer 22

Dimethyl sulfoxide - Tissue Culture Grade; cryoprotectant

Question 23

Reagent (Cell Culture): Ethanol

Answer 23

Ethanol (70%) - Used to decontaminate

Question 24

Reagent (Cell Culture): Isopropanol

Answer 24

Isopropanol (100%)

Question 25

Reagent (Colony Forming Units): Crystal Violet

Answer 25

Crystal Violet

Question 26

Reagent (Colony Forming Units): Methanol

Answer 26

Methanol (100%)

Question 27

Supplies (Cell Culture): Sterile plastic disposable serological pipettes: 5ml, 10ml, 25ml, 50ml

Answer 27

Used when transferring solutions via aspiration. Located under the hood on the shelves below the top. Always open from the thicker side.

Question 28

Supplies (Cell Culture): Sterile plastic disposable pipettes for vacuum aspiration

Answer 28

Used when discarding solutions via aspiration. Located under the hood on the top shelf. Always open from the thicker side.

Question 29

Supplies (Cell Culture): Sterile plastic disposable serological pipettes: 5ml, 10ml, 25ml, 50ml ????

Answer 29

Used when transferring solutions and serums via aspiration or when "mixing" compact centrifuged cells back into medium. Located under the hood on the shelves below the top. Always open from the thicker side. ????

Question 30

Supplies (Cell Culture): Sterile plastic disposable pipettes for vacuum aspiration

Answer 30

Used when discarding solutions via vacuum aspiration. Located under the hood on the top shelf. Always open from the thicker side.

Question 31

Supplies (Cell Culture): Sterile plastic disposable conical centrifuge tubes

Answer 31

Coming in at 15ml and 50ml, these tubes are placed in the centrifuge to bring the cells to the bottom. Be sure to always use a counterweight when operating the centrifuge.

Question 32

Supplies (Cell Culture): Sterile plastic disposable serological pipettes: 5ml, 10ml, 25ml, 50ml ????

Answer 32

Located under the hood on the shelves below the top. Always open from the thicker side. ????

Question 33

Supplies (Cell Culture): Sterile plastic disposable pipettes for vacuum aspiration

Answer 33

Used when discarding solutions via vacuum aspiration. Located under the hood on the top shelf. Always open from the thicker side.

Question 34

Supplies (Cell Culture): Automated pipetter

Answer 34

This device accurately measures from 10 to 1000 microliters

Question 35

Supplies (Cell Culture): Sterile plastic disposable conical centrifuge tubes

Answer 35

Coming in at 15ml and 50ml, these tubes are placed in the centrifuge to bring the cells to the bottom. Be sure to always use a counterweight when operating the centrifuge.

Question 36

Supplies (Cell Culture): Sterile Tissue Flasks

Answer 36

Used to stored the cells in the incubator, breathable.

Question 37

Supplies (Cell Culture): Cryo-vials

Answer 37

2.0ml, used to freeze cells down to -180 degrees Celsius

Question 38

Supplies (Cell Culture): Cryo-freezing container

Answer 38

This contains 100% isopropanol.

Question 39

Supplies (Differentiation Assays): Costar 6 well TC treated microplates

Answer 39

Costar 6 well TC treated microplates

Question 40

Equipment: Laminar Flow Hood

Answer 40

Open it up and get to work!

Question 41

Equipment: Waterbath

Answer 41

37 degrees C and filthy from warm, sitting water

Question 42

Equipment: Tissue Culture Incubator

Answer 42

Located in the back room, gas and temperature controlled, if the alarm is sounding make sure to immediately notify a supervisor

Question 43

Equipment: Vacuum Aspiration Source

Answer 43

Plastic tube. Make sure to spray with alcohol after every use

Question 44

Equipment: Hemocytometer with coverslips

Answer 44

a counting-chamber device originally designed and usually used for counting blood cells. The hemocytometer was invented by Louis-Charles Malassez and consists of a thick glass microscope slide with a rectangular indentation that creates a chamber.