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Effects of glucose concentration in the medium on rat hepatocyte culture.

Na GH, Kim DG, Kim YH, Han JH, Jung ES - Ann Surg Treat Res (2014)

Bottom Line: Total cell count and viability showed smaller increases in the low-glucose group than the high-glucose group, although the difference was not statistically significant (P = 0.112 and P = 0.147, respectively).The levels of albumin (P = 0.943), ammonia (P = 0.744), and urea (P = 0.709) were not significantly different between the two groups.In both groups, the function of cultured hepatocytes decreased significantly over time.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Seoul St. Mary's Hospital, The Catholic University of Korea College of Medicine, Seoul, Korea.

ABSTRACT

Purpose: To determine the optimum culture conditions by investigating isolated rat hepatocytes cultured in medium containing different glucose concentrations.

Methods: Hepatocytes were isolated from rats using a two-step perfusion technique and divided into the following two groups cultured in medium containing different glucose concentrations: (1) low-glucose group and (2) high-glucose group. Total cell count and viability of cultured rat hepatocytes and liver function parameters (i.e., concentrations of albumin, ammonia, and urea in the culture medium) were measured. The morphology of cultured rat hepatocytes was examined by staining with hematoxylin and eosin, and albumin receptor expression was confirmed by immunofluorescence.

Results: Total cell count and viability showed smaller increases in the low-glucose group than the high-glucose group, although the difference was not statistically significant (P = 0.112 and P = 0.147, respectively). The levels of albumin (P = 0.943), ammonia (P = 0.744), and urea (P = 0.709) were not significantly different between the two groups. In both groups, the function of cultured hepatocytes decreased significantly over time. The morphology of hepatocytes was well maintained in both groups at 3 days. On day 7, the cytoplasm was transformed into a spindle shape. On day 10, these changes were exaggerated, and were more prominent in the high-glucose group.

Conclusion: Morphological assessment indicated that low-glucose culture medium is better than high-glucose culture medium for culturing of hepatocytes, although there was not significantly different in functional assessment. The cultured hepatocytes with low-glucose culture medium could be maintained for 7 days.

No MeSH data available.


Related in: MedlinePlus

Operative view of cannulation in the portal vein (A) and infusion of collagenase solution with whitish discoloration of the liver (B).
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Figure 1: Operative view of cannulation in the portal vein (A) and infusion of collagenase solution with whitish discoloration of the liver (B).

Mentions: The two-step perfusion method described by Seglen [7] was employed. First, Ca2+-free medium (Gibco 17701, Life Technologies, Grand Island, NY, USA) was perfused through the portal vein at a rate of 25 mL/min for 10 minutes. Second, Ca2+ and collagenase medium (Gibco 17703, Life Technologies) was perfused through the same route at the same rate. The perfused rat liver was disconnected from the perfusion apparatus and resected (Fig. 1).


Effects of glucose concentration in the medium on rat hepatocyte culture.

Na GH, Kim DG, Kim YH, Han JH, Jung ES - Ann Surg Treat Res (2014)

Operative view of cannulation in the portal vein (A) and infusion of collagenase solution with whitish discoloration of the liver (B).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4127907&req=5

Figure 1: Operative view of cannulation in the portal vein (A) and infusion of collagenase solution with whitish discoloration of the liver (B).
Mentions: The two-step perfusion method described by Seglen [7] was employed. First, Ca2+-free medium (Gibco 17701, Life Technologies, Grand Island, NY, USA) was perfused through the portal vein at a rate of 25 mL/min for 10 minutes. Second, Ca2+ and collagenase medium (Gibco 17703, Life Technologies) was perfused through the same route at the same rate. The perfused rat liver was disconnected from the perfusion apparatus and resected (Fig. 1).

Bottom Line: Total cell count and viability showed smaller increases in the low-glucose group than the high-glucose group, although the difference was not statistically significant (P = 0.112 and P = 0.147, respectively).The levels of albumin (P = 0.943), ammonia (P = 0.744), and urea (P = 0.709) were not significantly different between the two groups.In both groups, the function of cultured hepatocytes decreased significantly over time.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Seoul St. Mary's Hospital, The Catholic University of Korea College of Medicine, Seoul, Korea.

ABSTRACT

Purpose: To determine the optimum culture conditions by investigating isolated rat hepatocytes cultured in medium containing different glucose concentrations.

Methods: Hepatocytes were isolated from rats using a two-step perfusion technique and divided into the following two groups cultured in medium containing different glucose concentrations: (1) low-glucose group and (2) high-glucose group. Total cell count and viability of cultured rat hepatocytes and liver function parameters (i.e., concentrations of albumin, ammonia, and urea in the culture medium) were measured. The morphology of cultured rat hepatocytes was examined by staining with hematoxylin and eosin, and albumin receptor expression was confirmed by immunofluorescence.

Results: Total cell count and viability showed smaller increases in the low-glucose group than the high-glucose group, although the difference was not statistically significant (P = 0.112 and P = 0.147, respectively). The levels of albumin (P = 0.943), ammonia (P = 0.744), and urea (P = 0.709) were not significantly different between the two groups. In both groups, the function of cultured hepatocytes decreased significantly over time. The morphology of hepatocytes was well maintained in both groups at 3 days. On day 7, the cytoplasm was transformed into a spindle shape. On day 10, these changes were exaggerated, and were more prominent in the high-glucose group.

Conclusion: Morphological assessment indicated that low-glucose culture medium is better than high-glucose culture medium for culturing of hepatocytes, although there was not significantly different in functional assessment. The cultured hepatocytes with low-glucose culture medium could be maintained for 7 days.

No MeSH data available.


Related in: MedlinePlus