KR19980703028A - Sertoli Cells as Neurorecovery Inducing Cells for Neurodegenerative Diseases - Google Patents

Abstract

The present invention relates to a method for producing a nutrient factor product of a normal material by implanting a Sertoli cell producing a nutrient factor of a normal material into tissue of a mammal in need of the nutrient factor.

Description

Sertoli Cells as Neurorecovery Inducing Cells for Neurodegenerative Diseases

FIELD OF THE INVENTION The present invention generally relates to cell transplantation and cell transplantation methods, in particular, to improve behavioral and functional disorders associated with neurological and neurodegenerative diseases after transplantation into the central nervous system (CNS).

[Background]

In the treatment of diseases such as, for example, wound healing, it is often more useful to treat areas of tissue damage, especially with local nutritional factors, than systemic treatment.

As another example, transplanting neural tissue into the central nervous system (CNS) of a mammal may include epilepsy, seizures, Huntington's disease, head injury, spinal injury, pain, Parkinson's disease, myelin deficiency, neuromuscular disorders, neuralgia, muscular dystrophy It is becoming an alternative treatment for neurological and neurodegenerative diseases including lateral sclerosis, Alzheimer's disease, and affective disorders of the brain. The incubation and clinical data suggest that it survives, integrates and provides functional recovery with transplanted cells (grafts) and host tissues used in cell transplant protocols for this type of neurodegenerative disease (see Sanberg et al. 1994).

The primary source of these grafts was the fetus. For example, fetal ventral midbrain tissue has proven to be a source of viable grafts for Parkinson's disease (see Lindvall et al. 1990: Bjoklund, 1992). Likewise, fetal striatum tissue has been successfully used as a graft material for Huntington's disease (see lsacson et al. 1986; Sanberg et al. 1994).

Animals with neurological function were transplanted into non-fetal cells and non-neuronal cells / tissues. For example, chromaffin cells obtained from adult donors were used for the treatment of Parkinson's disease. The main advantage of this type of transplant protocol is that there is no ethical and logical problem associated with obtaining fetal tissue since the source is not a fetal source. Using chromogenic affinity protocols, normalized behavior is observed. However, recovery of this behavioral function is temporary and the animal returns to its pre-transplantation state (see Bjorklund and Stenevi, 1985; Lindvall et al. 1987). In the Parkinson's disease model, protocols are inadequate to keep the animals active in normal behavior, so the clinical application of these protocols as well as other treatments are premature.

Administration of growth factors as a method of treating neurological and neurodegenerative diseases has been noted in the art. However, transporting these drugs to the brain still presents many difficulties in successfully overcoming them. In general, these drugs cannot be computerized and injection into the brain is an impractical and incomplete solution. Cell manipulations that transport one specific nutrient in brain transplantation have been suggested, but the survival and stable transfection of cells during brain transplantation is still questionable. In addition, there is increasing recognition that several nutritional factors that work in concert in successful treatment of neurological and neurodegenerative conditions will be needed.

Long-lasting functional recovery was observed in diabetic animal models using new treatment protocols with isolated islet and sertoli cells. It is evident that this therapeutic effect is due to the presence of sertoli cells, in part due to their known immunosuppressive secretion factors (Selawry and Cameron, 1993; 1990 by Cameron et al.). Sertoli cells are also known to secrete a number of important trophic growth factors.

Therefore, it would be desirable to use only Sertoli cells as a source for disease where growth and trophic factors that aid injured tissue are useful. Examples include neurological diseases and wound treatment, including neurodegenerative diseases. Sertoli cells can be used to act as a normal plant for nutritional factors to promote wound healing and ameliorate functional and behavioral disorders associated with neurological and neurodegenerative diseases.

[Summary of invention]

The present invention provides a method for providing a nutrient factor product of a normal material by implanting a Sertoli cell secreting a nutrient of the normal material into a mammal.

Other advantages of the present invention will be readily appreciated, and the following detailed description will be better understood upon reference to the accompanying drawings.

FIG. 1 is a graph showing the results of apomorphine-induced rotational motion of two groups of animals showing a total of 7 rotations per minute or 210 rotations (large to lesion) when challenged with apomorphine prior to transplantation. Animals administered with medium alone continued to show significant rotation after transplantation, while animals receiving Sertoli cells showed a significant decrease in rotational motion (above 60%) after transplantation;

FIG. 2 is a graph showing the biased swing behavior of two groups of animals with 80% biased swing activity (large to lesion) as indicated by elevated body swing experiments. While animals exhibited significantly biased swing activity, the sertoli cells did not show any biased swing behavior after transplantation;

3A-C are optical micrographs showing ventral midbrain (VM) cells of fetal rats isolated and cultured for 7 days in control medium (CM) or Sertoli cell pre-regulated medium (SCM) for interference contrast optics in the dark field. It was taken with. FIG. 3A shows VM cells cultured in CM that show no signs of stimulation or differentiation. FIG. 3B shows highly stimulated state as VM cells cultured in SCM, and FIG. 3C shows that Sertoli cells secrete nutrients. VM cells cultured in SCM with trophic growth are shown at high magnification;

FIG. 4A is an electron micrograph showing the brain striatum showing the site of transplantation of the Sertoli cells and the infiltration tube (arrow). FIG. 4B is an electron micrograph showing a high magnification and high resolution of the box region shown in FIG. 4A. (Arrows) are readily identified because of the 1 μlatex cell content added intracellularly prior to transplantation;

Figures 5A-B are two optical micrographs of normal material transplanted Sertoli cells labeled with fluorescent markers (DiI) prior to transplantation into the brain striatum. Figure 5A is a rat host that was not treated with cyclosporine A in immunosuppressive treatment. Viable fluorescent sertoli cells are shown, and FIG. 5B shows viable fluorescent sertoli cells in rat hosts subjected to cyclosporin A immunosuppressive treatment.

In general, the present invention provides a method for facilitating repair, protection and aid of dysfunctional tissues by mechanisms including normal material production of Sertoli cell-induced growth factors and regulatory factors, which are considered nutritional factors. In another aspect, the present invention provides a method for producing a nutrient factor product of the normal material. This is done by transplanting isolated Sertoli cells, which are cells that secrete normal nutrients, into mammals.

An important advantage of using Sertoli cells as a normal production plant for nutrient factor production is that Sertoli cells have been shown to have an efficient immunosuppressive effect. Thus, there is no need for ancillary treatment causing immunosuppression. That is, Sertoli cells can be used as a nutritional factor while providing local self-induced immunosuppressive effects.

Nutritional factors secreted by Sertoli cells include Sertoli cell-induced growth factors and regulatory factors such as insulin-like growth factors I and II, epithelial growth factors, transforming growth factors α and β, and interleukin 1α ( Griswold, 1992). The wider range of Sertoli cell secretion factors is shown in Table 1. These factors have been shown to improve the effects on behavioral and functional disorders associated with neurodegenerative diseases. These factors are well known as nutrients that aid normal cell and tissue metabolism and function (see Grisold, 1992). The present invention takes advantage of the phenomenon that Sertoli cells can produce a nutrient-rich growth-assisted fluid microenvironment at the site of cell dysfunction or cell / tissue damage. Cell / tissue damage includes, but is not limited to, radiation damage, burns, and wounds. In contrast to the Sertoli cell / island cell transplant protocol used in the diabetic model, the method of the present invention attempts to transplant two different cell types into one host site by using only one type of cell, ie, Sertoli cell. It can solve a lot of logical procedural problems.

In the examples below, rat Sertoli cells are used, but Sertoli cells from any suitable source can be used. For example, human Sertoli cells will be used for human transplantation. Further, in a preferred embodiment of the present invention, pig sertoli cells can be transplanted into a mammal, such as a human body. Moreover, the veterinary use of the present invention is noteworthy and heterologous sertoli cells will be selective for transplantation into a preferred mammalian host.

As described in the experimental section below, the present invention can be used as a therapy for ameliorating behavioral and functional disorders associated with neurodegenerative diseases such as Huntington's disease and Parkinson's disease. This can be done without the side effects of immunosuppressive adjuvant therapy such as the chronic use of cyclosporin A previously used. Sertoli cells provide both secretory and immunosuppressive effects of trophic factors.

As shown in the examples below, transplantation of Sertoli cells prior to brain lesion induction or formation may provide a neuroprotective effect. For example, transplanting Sertoli cells prior to the induction of Huntington's disease, as described below, has both neuroprotective and prophylactic effects on subsequent brain lesions. Therefore, implanting Sertoli cells early after diagnosis of neurodegenerative disease would be useful for the treatment, prevention or alleviation of the disease. Sertoli cells can also be implanted for other types of CNS trauma, such as head injury, to mitigate the effects of CNS damage to the therapeutic, prophylactic and / or prophylactic level.

The following examples illustrate the ability of the present invention to ameliorate behavioral disorders associated with neurodegenerative diseases.

Example 1 Sertoli Cell Transplantation

Specific protocol:

The protocol generally involves two basic steps: (1) isolation of Sertoli cells and (2) cell transplantation, both of which are briefly described below and described in more detail by Selawry and Cameron (1993). See Pakzaben et al. (1993) for cell transplantation. Both documents are incorporated herein by reference.

(1A) Isolation of Sertoli Cells

The separation process follows the well-defined methods of Selawry and Cameron (1993) and is typically used. The cell culture medium used for all separation steps and cell culture was FMEM of DMEM: ham supplemented with retinol, ITS and gentamicin sulfate (Cameron and Muffily, 1991). The testicles of 16-day-old male Sprague-Doleley rats were surgically taken. In order to separate other testicular cell types from Sertoli cells, testicles were encapsulated to prepare enzymatic incisions. Enzymatic procedures used collagenase (0.1%), hyaluronidase (0.1%), and trypsin (0.25%), typical procedures used in many cell separation protocols. After serial enzymatic dissection, Sertoli cell isolates were washed with culture medium, transferred to sterile incubator and placed in a 5% CO ₂ -95% air humidified tissue culture incubator. After 48 hours incubation in a 39 ° C. incubator in advance, the Sertoli cells were washed to remove all contaminants. The resulting Sertoli cell-rich fractions were resuspended in 0.25 ml of DMEM / F12 medium and incubated at 37 ° C. for at least 24 hours.

Sertoli cells were liberated with trypsin at the bottom of the vessel and transferred to sterile conical test tubes, followed by washing by centrifugation and treatment with trypsin inhibitor to stop the enzyme action of trypsin. During the implantation period, the Sertoli cell-rich fraction was resuspended and sucked using a 20 gauge needle with a Hamilton syringe.

(1B) Isolation and Pretreatment of Sertoli Cells

Alternatively, as described above (1987a by Cameron et al .; 1987b by Cameron et al.) Testicles of the anesthetized rat using 0.25% trypsin (purchased from Sigma) and 0.1% collagenase (purchased from Sigma, type V) Was enzymatically processed at 37 ° C. (see Cameron et al. 1987a; Cameron et al. 1987b). The resulting Sertoli cell aggregates were evenly distributed in a 20 ml volume culture medium in a 75 cm 2 tissue culture flask (purchased from Costa). Cells were stored in sterilized 0.5 mM Tris-HCl buffer for 1 minute to remove contaminated germ cells (see Galdieri et al. 1981), and then the plated Sertoli aggregates were treated with 5% CO ₂ -95% for 48 hours. Incubated at 39 ° C. in air. After washing twice with culture medium, the flask was filled with 20 ml of culture medium and returned to 37% 5% CO ₂ -95% air 5% CO ₂ -injected thermostat. The resulting pretreated Sertoli-rich culture contained 95% or more Sertoli cells. Plate density (2.0 × 10 ⁶ Sertoli / cm 2) was not the result of frontal tomography of the cells.

(2) cell transplantation

The transplant protocol follows the procedure previously described in Pakzaban et al. 1993. Surgery of the animals was performed under sterile conditions. Initially all animals were anesthetized with 0.60 ml / kg of pentobarbital sodium and placed in a Koph stereotaxic instrument. Unilateral striatum grafts were performed in equivalent sets: anteroposterior = + 1.2, mesolateral = + / − 2.8, ventral direction = 6.0, 5.9 and 5.8 (based on the diagram of Paxinos and Watson, 1984). The striatum ipsilateral to the damaged black matter was implanted into Sertoli cells. A total of 3 μl volume of Sertoli cell suspension was added to each striatum. 1 μl of Sertoli cell suspension was injected at the abdominal site for at least 1 minute. Only medium was added to the control. Allow another 5 minutes to reach the final abdominal area before removing the needle. After surgery, animals were placed on an electric blanket to recover. Cyclosporin-A (20 mg / kg / day, intraperitoneal) was used immediately after surgery and on the day of implantation to provide short-term immunosuppression. Ongoing studies, however, have proven that such short-term cyclosporin-A is unnecessary (see also 5A-B).

Sertoli cells are transplanted into animal models of various neurodegenerative diseases by stereoposition equally defined for a particular disease, as described, for example, in Parkinson's disease, and systemic assays for functional recovery by techniques specific to that animal model This is done.

This study used 8-week-old male Sprag-Doleley rats with 6-OHDA-induced paraplegia (n = 12). Three weeks after the lesion, animal behavioral experiments were conducted, including apomorphine-induced rotational motion and swing motion. Baseline data showed significant apomorphine-induced rotational behavior (contrast to the lesion side of the CNS) in all these animals (at least 200 times for 30 minutes). In addition, significant right-biased swing activity (more than 70%) was recorded using the elevated body swing test (EBST).

Three weeks after the lesion, the same surgery was performed except that one group of animals (n = 6) received sertoli cells and one group received only medium (DMEM without serum) as a control. All animals received cyclosporin (20 mg / kg) the first two days after transplantation. One month, one and a half months and two months after transplantation, the same behavioral experiments were repeated for animals.

Animals that received sertoli cells showed a significant decrease in rotation (average 50 times in 30 minutes), whereas animals that received medium alone had rotational levels before transplantation (see Figure 1). Normalization of rotational motion continued over the course of two months of experiments. The postal swing activity shown previously by Sertoli cell transplanted animals was also significantly reduced during the post-transplantation experiment (see Figure 2). Animals treated with medium did not show a significant decrease in postal swing response.

Upon dissection, the brains of the animals were removed and fixed for vibratom sectioning by 40-80 μm. After staining, the glial cells activated at the infiltration site (ie, the lesion site) of the Sertoli cell transplanted rats were markedly reduced when comparing only the infiltration sites in the lesion animals without transplantation of the Sertoli cells.

Example 2: Growth of Neurons

Culture medium and Sertoli cell pre-conditioning medium

The culture medium used for Sertoli cell culture and co-culture is Dulbecco's minimum essential medium: 1: 1 F12 nutrient medium from Ham (available from Weater Bioproducts), 3 mg / ml L-glutamine (from Sigma) , Grade III), 0.01 cc / ml insulin-transferrin-selenium (ITS, purchased from Collaborative Research, Inc.), 50 ng / ml retinol (purchased from Sigma), 19 μl / ml lactic acid (from Sigma) Purchased) and 0.01 cc / ml gentamycin sulfate (obtained from Gibco).

After the initial 48 hours of culture of isolated Sertoli cells, the media were collected and centrifuged at 1500 rpm for 5 minutes. Supernatants were collected and immediately frozen in sterile test tubes. This medium was considered Sertoli pre-conditioned medium (SCM).

Isolation and Culture of Fetal Brain Cells

Fetal brain cells (FBCs) were collected from the ventral midbrain of fetal rats (15-17 days of pregnancy). Fetal brain tissue was initially dispersed by a series of subcutaneous needles (18-26 gauges) suspended in media and successively decreasing in size. The resulting suspension was treated with 0.1% trypsin for 5 minutes and then treated with 0.1% trypsin inhibitor for 2 minutes. Suspended FBS was washed three times, resuspended in culture medium and plated in a poly-L-lysine-coated incubator.

The ventral midbrain (VM) cells of fetal rats were isolated and incubated for 7 days in control medium (CM) or Sertoli cell pre-regulated medium (SCM) as shown in FIG. 3A. VM cells cultured in CM showed no signs of cell stimulation or differentiation. Referring to FIG. 3B, VM cells cultured in SCM were highly stimulated. Figure 3C shows a high magnification of the nutrient growth of the VM cells cultured in SCM in response to Sertoli cell secretion nutrient factors.

Example 3: Identification of Sertoli Cells

Insertion of latex beads:

Sertoli cells were isolated and cultured as described. Prior to implantation (approximately 12 hours), sterile 1 μm latex stock (10 μl / ml medium; purchased from Pelco, Tustin, Calif.) Was added to the culture medium. Sertoli cells rapidly phagocytized the beads. Immediately before transplantation, the columnar Sertoli cells were washed three times and resuspended in 1 ml of culture medium.

In Figure 4A, Sertoli cells were implanted into the brain's striatum, where the infiltration tract (arrow) and the Sertoli cell transplant site are shown. As shown in FIG. 4B, at high magnification, the sertoli cells (arrows) were easily identified because they contained 1 µ latex strains loaded into the sertoli cells before transplantation.

Example 4 Effect of Cyclosporin A (CAS) on Survival of Transplanted Sertoli Cells]

Fluorescent cell markers:

Just prior to implantation (approximately 2 hours) at 37 ° C. with CM-DiI fluorescent dye (100 μl stock solution / ml medium; purchased from Molecular Probes, Eugene, Oregon, Inc.) for cell tracking After 7 minutes of treatment, the solution was further placed at 4 ° C for 15 minutes. Fluorescent signs Serpoli cells were washed three times and resuspended in 1 ml of culture medium.

The effect of cyclosporin A on the survival of normal material transplanted sertoli cells was examined. Prior to transplantation into the striatum of the brain, transplanted Sertoli cells were labeled with fluorescent markers (DiI). One month after transplantation, tissues were collected. In FIG. 5A, fluorescent sertoli cells viable in rat hosts that were not immunosuppressed with cyclosporin A were observed. In Figure 5B, viable fluorescent sertoli cells are observed in rat hosts not subjected to cyclosporin A immunosuppressive treatment. This experiment demonstrates that cyclosporin A is not required for the survival of Sertoli cells transplanted into the brain.

Example 5 Preventive Effect of Sertoli Cells

Transplantation of Sertoli cells is neuroprotective when transplanted prior to induction of brain lesions. The protective effect of these Sertoli cells has been demonstrated in animal models for Huntington's disease (HD). This model is produced by systemic administration of the mitochondrial inhibitor 3-nitroproproic acid (3NP). Sanberg and his colleagues (Koutouzis et al. 1994; Borlongan et al. 1995) and others have demonstrated that 3NP infusion causes specific lesions similar to those seen in Huntington's disease in the striatum.

In this experiment, 8 rats were implanted with sertoli cells of a rat to one side of a normal striatum of the rat (as described above). Thus, there was a Sertoli cell on one side of the brain and no on the other.

One month later, HD was induced by injecting 3NP into the animal as described in the literature (Koutouzis et al. 1994; Borlongan et al. 1995). Upon injection of 3NP, normal rats are injured on both sides of the brain striatum and have behavioral disorders equally on both sides of the body (see Koutouzis et al. 1994; borlongan et al. 1995).

One month after 3NP administration, the animals showed unilateral behavioral disorders. This was observed by post-lesion apomorphine-induced rotation experiments in sertoli transplanted animals, not in controls (number of rotations: control = 0.25 ± 6; sertoli transplant = 197 ± 31.9, P.0001). This asymmetric rotational motion suggested a lesion in the brain on which the Sertoli cells were not implanted. Thus, Sertoli cell transplantation for nutritional mechanisms has neuroprotective and prophylactic effects on subsequent brain lesions. This provides evidence that Sertoli transplantation is also useful for early treatment of neurodegenerative diseases before they are severely impaired.

These results, taken together, show that Sertoli cells ameliorate behavioral and functional deficiencies in Parkinson's and Huntington's disease animal models. A related mechanism would be the secretion of serotoli cell-induced growth factors and regulatory factors that promote the sustained support and recovery of related neural tissues, as described by the growth of neuronal tissue in Example 2. Sertoli cells can also protect and promote neuronal tissue recovery in the brain by inhibiting glial cell activation at the lesion site. These results also demonstrate the viability of normal material transplanted sertoli cells.

Throughout this application, many publications are referred to by quotation or number. Complete citations for the publications are listed below. To more fully describe the technical aspects pertaining to this application, the entirety of these publications is incorporated herein by reference.

The present invention has been described in an illustrative format, and it is to be understood that the terminology which has been used is intended to be in the nature of words of description rather than of limitation.

Obviously, various modifications and variations of the present invention are possible in light of the above guidelines. Accordingly, it is recognized that, within the scope of the appended claims, the invention may be practiced otherwise than as detailed.

TABLE 1

I. Sertoli Cell-Induced Growth Factors and Regulators (Partial List)

Category and Protein Function

Hormone / growth factor inhibition

Miller's Inhibitors

Inhibition of Inhibin FSH Release

Insulin-like growth factor

(Somatomedin A and C, IGF) Growth Factor

Prodinorphine

Interleukin-1α mitogen

Transformation Growth Factor αβ Growth Factor

Basic fibroblast growth factor growth factor

LHRH-Like Factor Leithi Cell Steroid Production

(Unrefined or incompletely specified)

Sertoli Secretion Growth Factor Growth Factor

Semi-production growth factor

Reich's cell stimulation activity

Testines

CMB Protein

Vitamin-binding protein vitamin transport

Transport and biological defense

Transfer iron transport

Celluloplasmic copper transport

Binding to Safosin Glycosphingolipids

SGP-2 (CLUSTERIN) Lipid Transport?

Androgen binding protein T and DHT transport

SPARC calcium binding protein?

IGF binding protein IGF transport

Riboflavin-binding Protein Riboflavin Transport

Proteases and Protease Inhibitors

Plasminogen activator protease

Cyclic Protein-2 Protease Inhibitor

Cystatin Protease Inhibitor

α ₂ -macroglobulin protease inhibitors

Collagen collagenase protease

Metalloproteinase proteases

Element

Collagen Ⅳ

Laminin

Proteoglycan

Claims (20)

A method of producing a nutritional factor of normal material by implanting a Sertoli cell producing a nutritional factor of normal material into the tissue of a mammal that requires the nutritional factor. The method of claim 1, wherein the tissue requiring nutritional factors is the central nervous system of a mammal. The method of claim 2, wherein the mammal suffers from a neurological disease, including neurodegenerative disease, the method further comprising ameliorating the behavioral and dysfunction caused by the disease by the action of secreted nutritional factors. . The method of claim 1, wherein the Sertoli cells are porcine Sertoli cells. The method of claim 2, wherein said transplanting step is further limited to protecting the central nervous system from degenerative diseases. 3. The method of claim 2, wherein said transplanting step is further limited to restoring damaged central nervous system tissue. The neurological or neurodegenerative disorders of claim 3 include epilepsy, seizures, Huntington's disease, head injuries, spinal cord injuries, pain, Pickinson's disease, myelin deficiency, neuromuscular disease, neuralgia, muscular dystrophy, Alzheimer's disease and brain Production method characterized by including the affective disorder of. To treat neurological diseases including epilepsy, Huntington's disease, head injury, spinal injury, pain, pain, Parkinson's disease, myelin deficiency, neuromuscular disease, neuralgia, amyotrophic lateral sclerosis, Alzheimer's disease and brain affective disorders A method of producing nutrient product of normal material by transplanting porcine Sertoli cells secreting nutrient of normal material into nervous system. The production method according to claim 8, wherein the test subject is a human body. A method of producing a nutrient product of a normal material by implanting a Sertoli cell that secretes a nutrient of the normal material into the tissue damage area of the subject. Use of Sertoli Cells to Produce Nutrient Factors in Normal Materials by Transplanting Sertoli Cells Producing Normal Factors into Tissues in Need of Nutrient Factors in Mammals. The use of Sertoli cells according to claim 1, wherein the tissue in need of nutritional factors is the central nervous system of a mammal. The use of claim 2, wherein the mammal suffers from a neurological disease, including neurodegenerative diseases, wherein said use improves the behavioral and dysfunction caused by the disease by the action of secreted nutritional factors. Use according to claim 1, wherein the Sertoli cells are porcine Sertoli cells. 3. Use of Sertoli cells according to claim 2, characterized in that the transplant is further limited to protecting the central nervous system from degenerative diseases. 3. The use of Sertoli cells according to claim 2, wherein said transplantation is further limited to restoring damaged central nervous system tissue. The neurological or neurodegenerative disorders of claim 3 include epilepsy, seizures, Huntington's disease, head injury, spinal cord injury, pain, Parkinson's disease, myelin deficiency, neuromuscular disease, neuralgia, muscular dystrophy, Alzheimer's disease and brain Use of Sertoli cells, characterized in that it comprises an affective disorder of. To treat neurological diseases including epilepsy, seizures, Huntington's disease, head injuries, spinal injuries, pain, Parkinson's disease, myelin deficiency, neuromuscular disease, neuralgia, Amyotrophic scleroderma, Alzheimer's disease and affective disorders of the brain, A sertoli cell useful for producing a trophic product of a normal material by transplanting porcine sertoli cells separating a nutrient of a normal material into the central nervous system of a subject. 19. The sertoli cell of claim 18, wherein the test subject is a human body. A sertoli cell useful for producing a nutrient product of a normal material by implanting a serotoli cell that secretes a nutrient of normal material into a tissue damage area of a subject.