Cell culture on semi-permeable tubular membranes

• US 3,883,393 A
• Filed: 02/11/1974
• Issued: 05/13/1975
• Est. Priority Date: 05/18/1972
• Status: Expired due to Term

• Current Assignee: Pietro M. Gullino, Richard A. Knazek, Robert L. Dedrick, William R. Kidwell
• Original Assignee: Pietro M. Gullino, Richard A. Knazek, Robert L. Dedrick, William R. Kidwell
• Inventors: Gullino, Pietro M., Knazek, Richard A., Dedrick, Robert L., Kidwell, William R.
• Primary Examiner(s): Rosen, Sam
• Application Number:
  US05/441,445
• Time in Patent Office: 456 Days
• Field of Search: 195/1.1,1.7,1.8,109,127,139 210/22,321
• US Class Current: 435/400
• CPC Class Codes: C12M 25/10 Hollow fibers or tubes hollow fiber modules in general B01D63/02
• CPC Class Codes: C12M 29/10 Perfusion
• CPC Class Codes: C12M 29/16 Hollow fibers hollow fiber modules in general B01D63/02

Abstract

The growth of cells in vitro on semi-permeable, tube-shaped membranes or capillaries is disclosed. Cells are initially allowed to settle onto the outer surfaces of the membranes in a nutrient medium environment while the membranes are continuously perfused by oxygenated nutrient medium flowing through the membranes. Nutrient substances diffuse from the perfusing medium through the membrane wall and into the cells, while cell products diffuse from the cell through the membrane wall into the perfusate from which the cell products may be recovered.

Claims

1. A METHOD FOR THE FORMATION AND MAINTANENCE OF SOLID TISSUES IN VITRO COMPRISING:

A. ARRANGING A MULTIPLICITY OF CAPILLARIES WITHIN A CHAMBER SIMULATING A VASCULAR NETWORK, THE CAPILLARIES HAVING WALLS WHICH ARE PERMEABLE TO NUTRIENTS REQUIRED FOR CELL GROWTH AND/OR CELL PRODUCTS AND BEING ARRANGED WITH INDIVIDUAL CAPILLARIES EXTENDING IN SUBSTANTIALLY PARALLEL RELTIONSHIP TO EACH OTHER WITHIN THE CHAMBER, DIVIDING THE CHAMBER BY THE WALLS OF THE CAPILLARIES INTO AN INTRACAPILLARY SPACE WITHIN THE CAPILLARIES AN EXTRAPILLARY SPACE OUTSIDE THE CAPILLARIES, THE INTRACAPILLARY SPACE AND THE EXTRACAPILLARY SPACE COMMUNICATING WITH EACH OTHER ONLY THROUGH THE WALLS OF THE CAPILLARIES, THE CAPOLLARIES BEING SPACED FROM EACH OTHER SO AS TO PROVIDE SUFFICIENT EXTRACAPILLARY SPACE FOR THREE-DIMENSIONAL GROWTH OF A LARGE NUMBER OF CELLS, WITH THE CAPILLARIES BEING IN SUFFICIENT PROXIMITY THAT WHEN THE DEPTH OF CELL GROWTH ON ONE CAPILLARY IS SUCH THAT THE CELLS GROWING ON THAT CAPILLARY FARTHEST FROM THAT CAPILLARY CAN NO LONGER OBTAIN NOURISHMENT FROM PERFUSATE PASSING THROUGH THAT CAPILLARY AND/OR REMOVAL OF WASTE PRODUCTS BY PERFUSATE PASSING THROUGH THAT CAPILLARY, SUCH CELLS WILL BE INFLUENCED BY PERFUSATE PASSING THROUGH AT LEAST ONE OTHER CAPILLARY, B. INTRODUCING LIVING CELLS INTO TGHE EXTRACAPILLARY SPACE SO THAT THE CELLS WILL SETTLE ONTO THE CAPILLARIES, AND C. PASSING PERFUSATE THROUGH THE INTRACPILLARY SPACE.

2. The method of claim 1 further including the step of recovering cell products passing from the cells through the capillary walls to the perfusate.

3. The method of claim 1 further including providing capillaries at least some of which have walls which are permeable to nutrients and/or cells products of large molecular weight and at least some of which have walls which are permeable to gases.

4. The method of claim 3 comprising providing capillaries having walls formed of different semi-permeable materials, some of which are more permeable to gases than others.

5. The method of claim 1 wherein the perfusate is oxygenated and pH controlled prior to being passed through the intracapillary space.