CHAPTER 2
Methods
A. M O S C O N A , O. A. T R O W E L L
and E. N. W I L L M E R
Department of Z°ol°gy> University of Chicago, U.S.A., M.R.C. Radiobiological
Unit, Harwell, England, and the Physiological Laboratory, University of
Cambridge, England.
I. Basic Requirements (E. N. Willmer)
9
A. Glassware
2
B. Water
2
99
C. Osmotic pressure
D. Hydrogen-ion concentration
3
E. Oxygen
5
F. Salt concentrations
^
97
G. Energy source
* '
H. Temperature
7
on
I. Size of tissue
o
I I . Tissue Culture (E. N. Willmer)
9
A. Hanging-drop method
9
B. Observation and perfusion chambers (slide chambers)
38
C. Flask cultures
4
D. Roller tubes
3
E. Tube cultures
4
I I I . Cell Culture
5
A. Cell isolation and pure strains of cells (E. N. Willmer)
45
B. Preparation of cell suspensions (A. Moscona)
9
C. Cell cultures (E. N. Willmer)
0
I V . Organ Culture (O. A. Trowell)
0
A. General comments
^
B. Organ fragments suspended in medium
72
C. Organs on a plasma clot
4
D. Organs on agar
j
E. Organs on a fluid medium
76
F. Perfusion methods
80
G. Limiting factors in organ culture
80
References
86
I. B A S I C R E Q U I R E M E N T S
AS suggested in the Introduction, the behaviour of animal* cells and
tissues in vitro very largely depends on the size of the units which are
*The methods for the study of plant tissues are discussed in Vol. 3, Chapter 9.