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Effect of medium osmolality on the growth of murine continuous marrow cultures
Abstract:
The effect of medium osmolality was examined in primary, continuous bone-marrow cultures established from TO strain mice. The non-adherent cell population increased exponentially between weeks 2 and 5 and thereafter declined steadily. The number of CFU-GM followed a similar pattern but showed greater variability. The optimum osmolality in 4 week old cultures was found to be about 345 mosmol/kg which was higher than the plasma osmolality (n = 20; mean = 323.3 mosmol/kg; range = 313-331). Maximum non-adherent cell numbers were found at about 345 mosmol/kg (better than half-maximum between 320 and 370 mosmol/kg). CFU-GM numbers in the culture supernatant were maximal at about 355 mosmol/kg (better than half-maximum between 320 and 400 mosmol/kg). An adherent layer developed over a wider range of osmolality than supported granulopoiesis (better than half-maximum between 258 and 402 mosmol/kg). It was necessary to increase the osmolality of Fischer's medium in order to obtain maximum growth.
Insights
Optimizing medium osmolality is crucial for bone marrow cultures. Higher osmolality than plasma, around 345-355 mosmol/kg, maximizes non-adherent cell and CFU-GM growth in mouse bone marrow cultures.
Area of Science:
- Hematology
- Cell Biology
- Biotechnology
Background:
- Primary bone marrow cultures are essential for studying hematopoiesis.
- Understanding optimal culture conditions, like medium osmolality, is key for reproducible results.
- Previous studies have not fully elucidated the impact of osmolality on mouse bone marrow cultures.
Purpose of the Study:
- To investigate the effect of varying medium osmolality on primary mouse bone marrow cultures.
- To determine the optimal osmolality for maximizing non-adherent cell proliferation and Colony-Forming Unit-Granulocyte-Macrophage (CFU-GM) counts.
- To compare the osmolality requirements for adherent layer formation versus granulopoiesis.
Main Methods:
- Primary bone marrow cells from TO strain mice were cultured continuously.
- Cultures were maintained under varying medium osmolality conditions.
- Non-adherent cell populations and CFU-GM numbers were quantified over time.
- Adherent layer development was also assessed across different osmolalities.
Main Results:
- Non-adherent cell populations and CFU-GM counts peaked at higher osmolalities (approximately 345-355 mosmol/kg) than physiological plasma osmolality (mean 323.3 mosmol/kg).
- Optimal osmolality for maximum non-adherent cell numbers was around 345 mosmol/kg, with a half-maximum range of 320-370 mosmol/kg.
- CFU-GM numbers were maximal at approximately 355 mosmol/kg (half-maximum range 320-400 mosmol/kg).
- Adherent layer formation occurred over a broader osmolality range (half-maximum 258-402 mosmol/kg) than granulopoiesis.
Conclusions:
- Medium osmolality significantly impacts mouse bone marrow culture outcomes, particularly for cell proliferation and CFU-GM production.
- An osmolality higher than physiological levels is required for optimal granulopoiesis in these cultures.
- Adjusting Fischer's medium osmolality is necessary to achieve maximal growth and cellularity in primary bone marrow cultures.