Kenichiro Asano

Formation of multicellular spheroids composed of adult rat hepatocytes in dishes with positively charged surfaces and under other nonadherent environments

Adult rat hepatocytes formed floating multicellular spheroids in primary culture in an uncoated plastic dish with a positively charged surface. Cells in the spheroids formed in such a simple way were similar to those formed in dishes coated with proteoglycan fraction isolated from rat liver reticulin fibers; in both cases, cells maintained high ability to produce albumin and poor ability to proliferate in response to epidermal growth factor. Coating dishes with albumin was also helpful in spheroid formation; coating with 2-hydroxymethyl methacrylate resulted in formation of incomplete spheroids. Elimination of serum factors was essential for the formation of spheroids; when cells were washed with serum-containing medium before seeding or if the medium was replaced with a serum-containing medium, spheroid formation was completely inhibited. Collagens, fibronectin, and laminin, all of which promote the adhesion and spreading of hepatocytes on substrates, inhibited spheroid formation. Furthermore, collagens disintegrated spheroids, and cells in the monolayer initiated proliferation. Thus, two distinct, mutually exclusive features of primary culture of adult hepatocytes apparently exist; monolayer culture with proliferative activity in an adherent environment and spheroid culture with poor proliferative activity and high albumin-producing ability in a nonadherent environment.

Promotion of Spheroid Assembly of Adult Rat Hepatocytes by Some Factor(s) Present in the Initial 6-Hour Conditioned Medium of the Primary Culture

Adult rat hepatocytes are capable of assembling to form floating multicellular spheroids (spheroids) in a dish with a positively charged surface in primary culture. In this report we show that the conditioned medium of the early time period of the culture exhibited biologically defined activity that promoted the assembly of isolated hepatocytes to floating spheroid. This activity was present only in the initial 6-hour conditioned medium; it was highest in the initial 2 h of culture and gradually decreased over 6 h and was not detected thereafter for 7 days. The conditioned medium appeared to inhibit the disintegration of spheroids that occurred during transfer to a new positively charged dish in the presence of either new or conditioned medium collected after day 4. Furthermore, disintegrated spheroids again assembled to form floating spheroid in the presence of the conditioned medium. Since the activity present in the conditioned medium was linearly dose-responsive to the inoculated viable cell number but not to that of dead cells, some factors responsible for the activity were probably produced by the viable cells during only a short time in culture.

The characteristics of patients with hypermagnesemia who underwent emergency hemodialysis

This study aimed to clarify the characteristics of patients who presented with severe hypermagnesemia and subsequently underwent emergency hemodialysis.

Impaired mental health status in patients with CKD is associated with eGFR decline: Lower MCS in CKD is associated with eGFR decline

Deteriorated health‐related quality of life (HRQOL) is associated with increased risk for death in both chronic kidney disease (CKD) and end‐stage renal disease (ESRD) patients; however, the impact of HRQOL on CKD progression is not well investigated.

Diffuse Alveolar Hemorrhage in IgA Vasculitis with an Atypical Presentation

IgA vasculitis (IgAV) commonly occurs in young children, who present with a tetrad of purpura, abdominal pain, arthralgia and nephritis. Diffuse alveolar hemorrhage (DAH) is a rare complication of IgAV. We herein report an adult case of IgAV with a presentation of DAH and nephritis (pulmonary renal syndrome, PRS), but without other typical manifestations, such as purpura, abdominal pain and arthralgia. A 33-year-old man presented with hemoptysis and a low-grade fever and was diagnosed to have IgAV based on the results of a renal biopsy. Treatment with corticosteroids, cyclophosphamide, and plasmapheresis was effective. IgAV should therefore be considered in the differential diagnosis of adult PRS.

A case of fatal osteolytic hypercalcemia complicated with IgG4-related ophthalmic disease leading to renal failure

A 40-year-old male was hospitalized with renal impairment and severe hypercalcemia. His concentration of serum IgG4 was high, but serum whole PTH, 1–25(OH)2 vitamin D3 and PTHrP were not elevated. Computed tomography showed swelling of the bilateral lacrimal glands and systemic lymphadenopathy. The histological findings of lacrimal gland biopsy fulfilled the diagnostic criteria of IgG4-related ophthalmic disease (IgG4ROD). Bone scintigraphy showed increased ectopic uptake in the stomach, heart, lungs, and kidneys. He died on day 16 of admission, although the therapies for hypercalcemia were continued. Autopsy results showed an increase of osteoclasts in the bone marrow and metastatic calcification in multiple organs, and excluded from the differential diagnosis other disorders which present lymph-node swelling and hypercalcemia such as cancer, lymphoma, Castleman’s disease, and sarcoidosis. He was given a diagnosis of IgG4ROD with osteolytic hypercalcemia.