Apparatus for mechanically stimulating cells

• US 4,851,354 A
• Filed: 12/07/1987
• Issued: 07/25/1989
• Est. Priority Date: 12/07/1987
• Status: Expired due to Fees

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Thibault, Lawrence E., Macarak, Edward J., Winston, Flaura K.
• Primary Examiner(s): Makay, Albert J.
• Assistant Examiner(s): KAMEN, NOAH P
• Application Number:
  US07/129,262
• Time in Patent Office: 596 Days
• Field of Search: 435/284, 435/285, 435/286, 435/296, 435/297, 435/298, 435/299, 215/6
• US Class Current: 435/402
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/22 Transparent or translucent parts glassware for laboratory use B01L3/00
• CPC Class Codes: C12M 23/38 Caps; Covers; Plugs; Pouring means
• CPC Class Codes: C12M 35/04 Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli

Abstract

An apparatus useful for studying cells in culture under conditions which reproduce their natural, in vivo mechanical environment is disclosed. The device comprises an airtight well having an optically transparent compliant base of a biologically compatible material on which said cells may be grown and an optically transparent, removable cap, coupled with a ported, airtight reservoir which reservoir has an optically transparent base and which reservoir can be filled with pressurizing media to create cyclic variations in hydrostatic pressure beneath said compliant base, causing said compliant base to deform and thereby exert a substantially uniform biaxial strain on the cells attached thereto.

Claims

1. A method for studying cells in culture under conditions which reproduce their natural, in vivo mechanical environment comprising

providing a device comprising

an airtight well having an optically transparent circular compliant base of a biologically compatible material on which said cells may be grown and an optically transparent, removable cap, which airtight well and removable cap are formed of biologically compatible materials, and

a ported, airtight reservoir coupled with said well beneath said compliant base, which reservoir has an optically transparent base;

seeding said cells on said compliant base;

allowing said cells to become confluent and attach to said compliant base;

injecting into and withdrawing from said airtight reservoir gaseous medium to create cyclic variations in hydrostatic pressure beneath said compliant base to concavedly-convexedly deform and thereby exert a substantially uniform biaxial strain on the cells attached thereto.

2. The method of claim 1 wherein said compliant base is a polyetherurethane urea film.

3. The method of claim 1 wherein said well, cap and reservoir are partially fabricated of stainless steel.

4. The method of claim 1 in which said compliant base is selected so that the biaxial strain exerted on said cells may be predicted from the hydrostatic pressure applied from below by the following equation:

space="preserve" listing-type="equation">biaxial strain=0.67(w.sub.o.sup.2 /a.sup.2)

where

a=inner radius of the well

w_o =(0.572)a[(P)a/Eh]^1/3

where

p=pressure applied from below

E=Young'"'"'s modulus

h=thickness of compliant base.

5. The method of claim 4 in which said compliant base exhibits substantially no auto-fluorescence.

6. The method of claim 4 in which said compliant base is a polyetherurethane urea film.

7. The method of claim 1 in which the gaseous medium is humidified air.

8. The method of claim 1 wherein said cells are endothelial cells.

9. The method of claim 8 wherein said cells are subjected to a cyclic biaxial strain of about 4.9% at 1 Hz.

10. The method of claim 1 in which said variations in hydrostatic pressure are wave shaped.