TECHNIQUE OF CULTIVATING HUMAN TISSUES IN VITRO

Area of Science:

• Tissue engineering
• Cell culture
• In vitro biology

Background:

• Unmodified human plasma is unsuitable for human tissue culture due to fibrin digestion by tissue ferments, hindering cell migration.
• Existing methods face challenges in maintaining tissue viability and framework integrity during culture.

Purpose of the Study:

• To develop an improved culture medium for sustained in vitro propagation of human tissues.
• To investigate the optimal conditions and limitations for human tissue culture.

Main Methods:

• Supplementing human plasma or serum with small amounts of fowl or pigeon plasma to enhance fibrin resistance.
• Culturing human tissues through serial subcultures over several months.
• Assessing tissue viability and growth after varying preservation periods and conditions.

Main Results:

• The addition of resistant avian plasma (fowl or pigeon) successfully prevented fibrin digestion, preserving the necessary framework for cell migration.
• Human tissues were propagated in the modified medium for extended periods (months) with potential for indefinite maintenance.
• Human tissues exhibited resilience to temperature changes and mechanical stress, comparable to lower animal tissues, and could be preserved for 6-8 days at 10-15°C.
• Autopsy-derived tissues showed active growth, indicating viability for several hours post-mortem.
• The presence of iso-antibodies in human serum did not affect tissue growth in vitro, negating any advantage of autologous serum.

Conclusions:

• A novel culture medium using resistant avian plasma significantly enhances the feasibility of long-term human tissue culture in vitro.
• Human tissues demonstrate robustness in culture, tolerating preservation and handling well, with potential for using autopsy-derived samples.
• Autologous serum offers no discernible benefit over homologous serum in human tissue culture systems.